def test_hello_world(self):
with tempdir.in_tempdir():
with tempfile.NamedTemporaryFile() as fasta:
with tempfile.NamedTemporaryFile() as tax:
fasta.write(Tests.extra_mcra_fasta)
fasta.flush()
tax.write(Tests.extra_mcra_taxonomy)
tax.flush()
prev_path = os.path.join(path_to_data,'mcrA.10seqs.gpkg')
cmd1 = "%s update --graftm_package %s --sequences %s --taxonomy %s --output %s" %(
path_to_script,
prev_path,
fasta.name,
tax.name,
'updated.gpkg')
extern.run(cmd1)
prev = GraftMPackage.acquire(prev_path)
up = GraftMPackage.acquire('updated.gpkg')
prevhash = prev.taxonomy_hash()
taxhash = up.taxonomy_hash()
self.assertEqual(len(prevhash)+1,
len(taxhash))
self.assertEqual(['mcrA','Euryarchaeota_mcrA','Methanofastidiosa'],
taxhash['KYC55281.1'])
self.assertEqual(prevhash['638165755'],
taxhash['638165755'])
seqio = SequenceIO()
self.assertEqual(
len(seqio.read_fasta_file(prev.unaligned_sequence_database_path()))+1,
len(seqio.read_fasta_file(up.unaligned_sequence_database_path())))
def test_hello_world(self):
with tempdir.in_tempdir():
with tempfile.NamedTemporaryFile() as fasta:
with tempfile.NamedTemporaryFile() as tax:
fasta.write(Tests.extra_mcra_fasta)
fasta.flush()
tax.write(Tests.extra_mcra_taxonomy)
tax.flush()
prev_path = os.path.join(path_to_data,'mcrA.10seqs.gpkg')
cmd1 = "%s update --graftm_package %s --sequences %s --taxonomy %s --output %s" %(
path_to_script,
prev_path,
fasta.name,
tax.name,
'updated.gpkg')
extern.run(cmd1)
prev = GraftMPackage.acquire(prev_path)
up = GraftMPackage.acquire('updated.gpkg')
prevhash = prev.taxonomy_hash()
taxhash = up.taxonomy_hash()
self.assertEqual(len(prevhash)+1,
len(taxhash))
self.assertEqual(['mcrA','Euryarchaeota_mcrA','Methanofastidiosa'],
taxhash['KYC55281.1'])
self.assertEqual(prevhash['638165755'],
taxhash['638165755'])
seqio = SequenceIO()
self.assertEqual(
len(seqio.read_fasta_file(prev.unaligned_sequence_database_path()))+1,
len(seqio.read_fasta_file(up.unaligned_sequence_database_path())))
def test_autodecorate(self):
with tempdir.in_tempdir():
with tempfile.NamedTemporaryFile() as fasta:
fasta.write(Tests.extra_mcra_fasta)
fasta.flush()
prev_path = os.path.join(path_to_data,'mcrA.10seqs.gpkg')
update = Update(prerequisites)
update.update(
input_sequence_path = fasta.name,
input_graftm_package_path = prev_path,
output_graftm_package_path = 'updated.gpkg')
prev = GraftMPackage.acquire(prev_path)
up = GraftMPackage.acquire('updated.gpkg')
prevhash = prev.taxonomy_hash()
taxhash = up.taxonomy_hash()
self.assertEqual(11, len(taxhash)) #hard-code 11 because of
#https://github.com/geronimp/graftM/issues/204
self.assertEqual(['mcrA','Euryarchaeota_mcrA', 'Methanomicrobia'],
taxhash['KYC55281.1'])
self.assertEqual(prevhash['638165755'],
taxhash['638165755'])
seqio = SequenceIO()
self.assertEqual(
len(seqio.read_fasta_file(prev.unaligned_sequence_database_path()))+1,
len(seqio.read_fasta_file(up.unaligned_sequence_database_path())))
def test_autodecorate(self):
with tempdir.in_tempdir():
with tempfile.NamedTemporaryFile() as fasta:
fasta.write(Tests.extra_mcra_fasta)
fasta.flush()
prev_path = os.path.join(path_to_data,'mcrA.10seqs.gpkg')
update = Update(prerequisites)
update.update(
input_sequence_path = fasta.name,
input_graftm_package_path = prev_path,
output_graftm_package_path = 'updated.gpkg')
prev = GraftMPackage.acquire(prev_path)
up = GraftMPackage.acquire('updated.gpkg')
prevhash = prev.taxonomy_hash()
taxhash = up.taxonomy_hash()
self.assertEqual(11, len(taxhash)) #hard-code 11 because of
#https://github.com/geronimp/graftM/issues/204
self.assertEqual(['mcrA','Euryarchaeota_mcrA', 'Methanomicrobia'],
taxhash['KYC55281.1'])
self.assertEqual(prevhash['638165755'],
taxhash['638165755'])
seqio = SequenceIO()
self.assertEqual(
len(seqio.read_fasta_file(prev.unaligned_sequence_database_path()))+1,
len(seqio.read_fasta_file(up.unaligned_sequence_database_path())))
def _assign_taxonomy(self, extracted_reads, assignment_method):
graftm_align_directory_base = os.path.join(self._working_directory,
'graftm_aligns')
os.mkdir(graftm_align_directory_base)
commands = []
all_tmp_files = []
# Run each one at a time serially so that the number of threads is
# respected, to save RAM as one DB needs to be loaded at once, and so
# fewer open files are needed, so that the open file count limit is
# eased.
for singlem_package, readsets in extracted_reads.each_package_wise():
tmp_files = []
for readset in readsets:
if len(readset.sequences) > 0:
tmp = tempfile.NamedTemporaryFile(prefix='singlem.%s' %
readset.sample_name,
suffix=".fasta")
# Record basename (remove .fasta) so that the graftm output
# file is recorded for later on in pipe.
tmpbase = os.path.basename(tmp.name[:-6])
readset.tmpfile_basename = tmpbase
seqio = SequenceIO()
seqio.write_fasta(readset.sequences, tmp)
tmp.flush()
tmp_files.append(tmp)
if len(tmp_files) > 0:
tmpnames = list([tg.name for tg in tmp_files])
cmd = "%s "\
"--threads %i "\
"--forward %s "\
"--graftm_package %s "\
"--output_directory %s/%s "\
"--max_samples_for_krona 0 "\
"--assignment_method %s" % (
self._graftm_command_prefix(singlem_package.is_protein_package()),
self._num_threads,
' '.join(tmpnames),
singlem_package.graftm_package_path(),
graftm_align_directory_base,
singlem_package.graftm_package_basename(),
assignment_method)
commands.append(cmd)
all_tmp_files.append(tmp_files)
extern.run_many(commands, num_threads=1)
for tmp_files in all_tmp_files:
[t.close() for t in tmp_files]
logging.info("Finished running taxonomic assignment with GraftM")
return SingleMPipeTaxonomicAssignmentResult(
graftm_align_directory_base)
def _align_sequences(self, input_sequences_path, output_alignment_path,
threads):
'''Align sequences into alignment_file
Parameters
----------
input_sequences_path: str
path to input sequences in fasta format
output_alignment_path: str
path to output alignment path
threads: str
number of threads to use
Returns
-------
Nothing
'''
logging.debug("Aligning sequences using mafft")
cmd = "mafft --anysymbol --thread %s --auto /dev/stdin > %s" % (
threads, output_alignment_path)
inputs = []
with open(input_sequences_path) as f:
for name, seq, _ in SequenceIO().each(f):
inputs.append('>%s' % name)
# Do not include * characters in the HMM, as this means tree
# insertion fails.
inputs.append(seq.replace('*', ''))
extern.run(cmd, stdin="\n".join(inputs))
def test_hello_world(self):
with tempfile.NamedTemporaryFile(prefix='graftm_decoy_test') as f1:
with tempfile.NamedTemporaryFile(prefix='graftm_decoy_test') as f2:
f1.write(self.eg1)
f1.flush()
extern.run("diamond makedb --in %s --db %s.dmnd" %\
(f1.name, f1.name))
f2.write(self.eg1)
f2.write(self.eg2)
f2.flush()
extern.run("diamond makedb --in %s --db %s.dmnd" %\
(f2.name, f2.name))
with tempfile.NamedTemporaryFile(
prefix='graftm_decoy_test') as f3:
with tempfile.NamedTemporaryFile(
prefix='graftm_decoy_test') as f4:
f3.write(self.eg1)
f3.flush()
ret = DecoyFilter(Diamond(f2.name + ".dmnd"),
Diamond(f1.name + ".dmnd")).filter(
f1.name, f4.name)
self.assertEqual(True, ret)
seqs = SequenceIO().read_fasta_file(f4.name)
self.assertEqual(1, len(seqs))
self.assertEqual("PROKKA_03952", seqs[0].name)
# clean up
os.remove(f1.name + ".dmnd")
os.remove(f2.name + ".dmnd")
def extract_and_read(self, reads_to_extract, database_fasta_file):
'''Extract the reads_to_extract from the database_fasta_file and return them.
Parameters
----------
reads_to_extract: Iterable of str
IDs of reads to be extracted
database_fasta_file: str
path the fasta file that containing the reads
Returns
-------
An array of graftm.sequence_io.Sequence objects'''
cmd = "fxtract -XH -f /dev/stdin '%s'" % database_fasta_file
process = subprocess.Popen(["bash", "-c", cmd],
stdin=subprocess.PIPE,
stdout=subprocess.PIPE)
output, error = process.communicate('\n'.join(reads_to_extract))
if process.returncode != 0:
raise Exception(
"Extraction command '%s' failed with exitstatus %i" %
(cmd, process.returncode))
seqs = []
for name, seq, _ in SequenceIO().each(StringIO(output)):
seqs.append(Sequence(name, seq))
return seqs
def _test_package(self, package_path):
'''Give a GraftM package a spin, and see if it works in reality with default
parameters (i.e. pplacer). If it does not work, then raise an error.
Parameters
----------
package_path: str
path to graftm_package to be tested
'''
pkg = GraftMPackage.acquire(package_path)
with tempdir.TempDir() as graftM_graft_test_dir_name:
# Take a subset of sequences for testing
with tempfile.NamedTemporaryFile(suffix=".fa", mode='w') as tf:
seqio = SequenceIO()
with open(pkg.unaligned_sequence_database_path()) as f:
seqio.write_fasta(
itertools.islice(seqio.each_sequence(f), 10), tf)
tf.flush()
cmd = "graftM graft --forward %s --graftm_package %s --output_directory %s --force" % (
tf.name, package_path, graftM_graft_test_dir_name)
extern.run(cmd)
def _test_package(self, package_path):
'''Give a GraftM package a spin, and see if it works in reality with default
parameters (i.e. pplacer). If it does not work, then raise an error.
Parameters
----------
package_path: str
path to graftm_package to be tested
'''
pkg = GraftMPackage.acquire(package_path)
with tempdir.TempDir() as graftM_graft_test_dir_name:
# Take a subset of sequences for testing
with tempfile.NamedTemporaryFile(suffix=".fa") as tf:
seqio = SequenceIO()
seqio.write_fasta(
itertools.islice(seqio.each_sequence(open(pkg.unaligned_sequence_database_path())), 10),
tf)
tf.flush()
cmd = "graftM graft --forward %s --graftm_package %s --output_directory %s --force" %(
tf.name, package_path, graftM_graft_test_dir_name)
extern.run(cmd)
def extract_and_read(self, reads_to_extract, database_fasta_file):
'''Extract the reads_to_extract from the database_fasta_file and return them.
Parameters
----------
reads_to_extract: Iterable of str
IDs of reads to be extracted
database_fasta_file: str
path the fasta file that containing the reads
Returns
-------
An array of graftm.sequence_io.Sequence objects'''
cmd = "mfqe --output-uncompressed --fasta-read-name-lists /dev/stdin --input-fasta '{}' --output-fasta-files /dev/stdout".format(
database_fasta_file)
# Retrieve each sequence exactly once so mfqe does not croak
output = extern.run(cmd, stdin='\n'.join(set(reads_to_extract)))
seqs = []
for name, seq, _ in SequenceIO().each(StringIO(output)):
seqs.append(Sequence(name, seq))
return seqs
def __init__(self):
self.clust = Deduplicator()
self.seqio = SequenceIO()
self.seq_library = {}
self.orfm_regex = OrfM.regular_expression()
class Clusterer:
def __init__(self):
self.clust = Deduplicator()
self.seqio = SequenceIO()
self.seq_library = {}
self.orfm_regex = OrfM.regular_expression()
def uncluster_annotations(self, input_annotations, reverse_pipe):
'''
Update the annotations hash provided by pplacer to include all
representatives within each cluster
Parameters
----------
input_annotations : hash
Classifications for each representative sequence of the clusters.
each key being the sequence name, and the entry being the taxonomy
string as a list.
reverse_pipe : bool
True/False, whether the reverse reads pipeline is being followed.
Returns
-------
output_annotations : hash
An updated version of the above, which includes all reads from
each cluster
'''
output_annotations = {}
for placed_alignment_file_path, clusters in self.seq_library.items():
if reverse_pipe and placed_alignment_file_path.endswith(
"_reverse_clustered.fa"):
continue
placed_alignment_file = os.path.basename(
placed_alignment_file_path)
cluster_classifications = input_annotations[placed_alignment_file]
if reverse_pipe:
placed_alignment_base = placed_alignment_file.replace(
'_forward_clustered.fa', '')
else:
placed_alignment_base = placed_alignment_file.replace(
'_clustered.fa', '')
output_annotations[placed_alignment_base] = {}
for rep_read_name, rep_read_taxonomy in cluster_classifications.items(
):
if reverse_pipe:
orfm_regex = OrfM.regular_expression()
clusters = {(orfm_regex.match(key).groups(0)[0]
if orfm_regex.match(key) else key): item
for key, item in iter(clusters.items())}
for read in clusters[rep_read_name]:
output_annotations[placed_alignment_base][
read.name] = rep_read_taxonomy
return output_annotations
def cluster(self, input_fasta_list, reverse_pipe):
'''
cluster - Clusters reads at 100% identity level and writes them to
file. Resets the input_fasta variable as the FASTA file containing the
clusters.
Parameters
----------
input_fasta_list : list
list of strings, each a path to input fasta files to be clustered.
reverse_pipe : bool
True/False, whether the reverse reads pipeline is being followed.
Returns
-------
output_fasta_list : list
list of strings, each a path to the output fasta file to which
clusters were written to.
'''
output_fasta_list = []
for input_fasta in input_fasta_list:
output_path = input_fasta.replace('_hits.aln.fa', '_clustered.fa')
cluster_dict = {}
logging.debug('Clustering reads')
if os.path.exists(input_fasta):
reads = self.seqio.read_fasta_file(
input_fasta) # Read in FASTA records
logging.debug('Found %i reads' %
len(reads)) # Report number found
clusters = self.clust.deduplicate(
reads) # Cluster redundant sequences
logging.debug('Clustered to %s groups' %
len(clusters)) # Report number of clusters
logging.debug(
'Writing representative sequences of each cluster to: %s' %
output_path) # Report the name of the file
else:
logging.debug("Found no reads to be clustered")
clusters = []
self.seqio.write_fasta_file(
[x[0] for x in clusters], output_path
) # Choose the first sequence to write to file as representative (all the same anyway)
for cluster in clusters:
cluster_dict[cluster[
0].name] = cluster # assign the cluster to the dictionary
self.seq_library[output_path] = cluster_dict
output_fasta_list.append(output_path)
return output_fasta_list
def __init__(self):
self.clust = Deduplicator()
self.seqio = SequenceIO()
self.seq_library = {}
self.orfm_regex = OrfM.regular_expression()
class Clusterer:
def __init__(self):
self.clust = Deduplicator()
self.seqio = SequenceIO()
self.seq_library = {}
self.orfm_regex = OrfM.regular_expression()
def uncluster_annotations(self, input_annotations, reverse_pipe):
'''
Update the annotations hash provided by pplacer to include all
representatives within each cluster
Parameters
----------
input_annotations : hash
Classifications for each representative sequence of the clusters.
each key being the sequence name, and the entry being the taxonomy
string as a list.
reverse_pipe : bool
True/False, whether the reverse reads pipeline is being followed.
Returns
-------
output_annotations : hash
An updated version of the above, which includes all reads from
each cluster
'''
output_annotations = {}
for placed_alignment_file_path, clusters in self.seq_library.iteritems():
if reverse_pipe and placed_alignment_file_path.endswith("_reverse_clustered.fa"): continue
placed_alignment_file = os.path.basename(placed_alignment_file_path)
cluster_classifications = input_annotations[placed_alignment_file]
if reverse_pipe:
placed_alignment_base = placed_alignment_file.replace('_forward_clustered.fa', '')
else:
placed_alignment_base = placed_alignment_file.replace('_clustered.fa', '')
output_annotations[placed_alignment_base] = {}
for rep_read_name, rep_read_taxonomy in cluster_classifications.iteritems():
if reverse_pipe:
orfm_regex = OrfM.regular_expression()
clusters={(orfm_regex.match(key).groups(0)[0] if orfm_regex.match(key) else key): item for key, item in clusters.iteritems()}
for read in clusters[rep_read_name]:
output_annotations[placed_alignment_base][read.name] = rep_read_taxonomy
return output_annotations
def cluster(self, input_fasta_list, reverse_pipe):
'''
cluster - Clusters reads at 100% identity level and writes them to
file. Resets the input_fasta variable as the FASTA file containing the
clusters.
Parameters
----------
input_fasta_list : list
list of strings, each a path to input fasta files to be clustered.
reverse_pipe : bool
True/False, whether the reverse reads pipeline is being followed.
Returns
-------
output_fasta_list : list
list of strings, each a path to the output fasta file to which
clusters were written to.
'''
output_fasta_list = []
for input_fasta in input_fasta_list:
output_path = input_fasta.replace('_hits.aln.fa', '_clustered.fa')
cluster_dict = {}
logging.debug('Clustering reads')
if os.path.exists(input_fasta):
reads=self.seqio.read_fasta_file(input_fasta) # Read in FASTA records
logging.debug('Found %i reads' % len(reads)) # Report number found
clusters=self.clust.deduplicate(reads) # Cluster redundant sequences
logging.debug('Clustered to %s groups' % len(clusters)) # Report number of clusters
logging.debug('Writing representative sequences of each cluster to: %s' % output_path) # Report the name of the file
else:
logging.debug("Found no reads to be clustered")
clusters = []
self.seqio.write_fasta_file(
[x[0] for x in clusters],
output_path
) # Choose the first sequence to write to file as representative (all the same anyway)
for cluster in clusters:
cluster_dict[cluster[0].name]=cluster # assign the cluster to the dictionary
self.seq_library[output_path]= cluster_dict
output_fasta_list.append(output_path)
return output_fasta_list
def generate_expand_search_database_from_contigs(self, contig_files,
output_database_file,
search_method):
'''Given a collection of search_hmm_files, search the contigs in
contig_files, and generate an HMM from the resulting hits, outputting
it as output_database_file.
Parameters
----------
contig_files: list of str
list of files to search
output_database_file: str
path to output file
search_method: str
"diamond" or "hmmsearch", to specify search method to use and what
type of database to build.
Returns
-------
True if genes were recovered, else False'''
ss = SequenceSearcher(self.search_hmm_files)
seqio = SequenceIO()
if search_method == self.DIAMOND_SEARCH_METHOD:
if self.diamond_database == None or self.unaligned_sequence_database == None:
logging.warning(
"Cannot expand_search continue with no diamond database or unaligned sequences."
)
return False
with tempfile.NamedTemporaryFile(
prefix='graftm_expand_search_orfs') as orfs:
logging.info("Finding expand_search hits in provided contigs..")
for contig_file in contig_files:
logging.debug("Finding expand_search hits in %s.." %
contig_file)
unpack = UnpackRawReads(contig_file)
with tempfile.NamedTemporaryFile(prefix='graftm_expand_search') as \
hit_reads_orfs_fasta:
# search and extract matching ORFs
with tempfile.NamedTemporaryFile(prefix='graftm_expand_search2') as \
hmmsearch_output_table:
with tempfile.NamedTemporaryFile(prefix='graftm_expand_search3') as \
hit_reads_fasta:
ss.search_and_extract_orfs_matching_protein_database(\
unpack,
search_method,
self.maximum_range,
self.threads,
self.evalue,
self.min_orf_length,
None,
(self.diamond_database if self.diamond_database else None),
hmmsearch_output_table.name,
hit_reads_fasta.name,
hit_reads_orfs_fasta.name)
# Append to the file
shutil.copyfileobj(open(hit_reads_orfs_fasta.name), orfs)
# Now have a fasta file of ORFs.
# Check to make sure the file is not zero-length
orfs.flush()
with tempfile.NamedTemporaryFile(
prefix="graftm_expand_search_aln") as aln:
if search_method == self.HMM_SEARCH_METHOD:
# Check that there is more than one sequence to align.
if len(
seqio.read_fasta_file(orfs.name)
) <= 1: # Just to build on this, you need to check if there is > 1 hit
# otherwise mafft will fail to align, causing a crash when hmmbuild is
# run on an empty file.
logging.warn(
"Failed to find two or more matching ORFs in the expand_search contigs"
)
return False
# Run mafft to align them
cmd = "mafft --auto %s >%s" % (orfs.name, aln.name)
logging.info("Aligning expand_search hits..")
extern.run(cmd)
# Run hmmbuild to create an HMM
cmd = "hmmbuild --amino %s %s >/dev/null" % (
output_database_file, aln.name)
logging.info("Building HMM from expand_search hits..")
extern.run(cmd)
elif search_method == self.DIAMOND_SEARCH_METHOD:
# Concatenate database with existing database
with tempfile.NamedTemporaryFile(
prefix="concatenated_database") as databasefile:
for f in [orfs.name, self.unaligned_sequence_database]:
for line in open(f):
databasefile.write(line)
databasefile.flush()
# Run diamond make to create a diamond database
cmd = "diamond makedb --in '%s' -d '%s'" % (
databasefile.name, output_database_file)
logging.info(
"Building a diamond database from expand_search hits.."
)
extern.run(cmd)
else:
raise Exception("Search method not recognised: %s" %
search_method)
return False
return True
else:
loglevel = logging.INFO
logging.basicConfig(level=loglevel,
format='%(asctime)s %(levelname)s: %(message)s',
datefmt='%m/%d/%Y %I:%M:%S %p')
# Read in taxonomy
logging.info("Reading taxonomy..")
gg = GreenGenesTaxonomy.read(open(args.greengenes_taxonomy)).taxonomy
logging.info("Read in %i taxonomies" % len(gg))
# Read in sequence
logging.info("Reading sequences..")
duplicates = set()
sequences = {}
for name, seq, _ in SequenceIO()._readfq(open(args.sequences)):
if name in sequences:
logging.error("Duplicate sequence name %s" % name)
duplicates.add(name)
else:
sequences[name] = seq
logging.warn("Found %i duplicated IDs" % len(duplicates))
for dup in duplicates:
del sequences[dup]
logging.info("Read in %i sequences" % len(sequences))
# Ensure that each sequence in the taxonomy has an associated sequence,
# otherwise delete it
tax_no_seq = set()
for name, taxonomy in gg.items():
if name not in sequences:
def main(self, **kwargs):
alignment = kwargs.pop('alignment',None)
sequences = kwargs.pop('sequences',None)
taxonomy = kwargs.pop('taxonomy',None)
rerooted_tree = kwargs.pop('rerooted_tree',None)
unrooted_tree = kwargs.pop('unrooted_tree',None)
tree_log = kwargs.pop('tree_log', None)
prefix = kwargs.pop('prefix', None)
rerooted_annotated_tree = kwargs.pop('rerooted_annotated_tree', None)
user_hmm = kwargs.pop('hmm', None)
search_hmm_files = kwargs.pop('search_hmm_files',None)
min_aligned_percent = kwargs.pop('min_aligned_percent',0.01)
taxtastic_taxonomy = kwargs.pop('taxtastic_taxonomy', None)
taxtastic_seqinfo = kwargs.pop('taxtastic_seqinfo', None)
force_overwrite = kwargs.pop('force',False)
graftm_package = kwargs.pop('graftm_package',False)
dereplication_level = kwargs.pop('dereplication_level',False)
threads = kwargs.pop('threads',5)
if len(kwargs) > 0:
raise Exception("Unexpected arguments detected: %s" % kwargs)
seqio = SequenceIO()
locus_name = (os.path.basename(sequences).split('.')[0]
if sequences
else os.path.basename(alignment).split('.')[0])
tmp = tempdir.TempDir()
base = os.path.join(tmp.name, locus_name)
insufficiently_aligned_sequences = [None]
removed_sequence_names = []
tempfiles_to_close = []
if prefix:
output_gpkg_path = prefix
else:
output_gpkg_path = "%s.gpkg" % locus_name
if os.path.exists(output_gpkg_path):
if force_overwrite:
logging.warn("Deleting previous directory %s" % output_gpkg_path)
shutil.rmtree(output_gpkg_path)
else:
raise Exception("Cowardly refusing to overwrite gpkg to already existing %s" % output_gpkg_path)
logging.info("Building gpkg for %s" % output_gpkg_path)
# Read in taxonomy somehow
gtns = Getaxnseq()
if rerooted_annotated_tree:
logging.info("Building seqinfo and taxonomy file from input annotated tree")
taxonomy_definition = TaxonomyExtractor().taxonomy_from_annotated_tree(\
Tree.get(path=rerooted_annotated_tree, schema='newick'))
elif taxonomy:
logging.info("Building seqinfo and taxonomy file from input taxonomy")
taxonomy_definition = GreenGenesTaxonomy.read_file(taxonomy).taxonomy
elif taxtastic_seqinfo and taxtastic_taxonomy:
logging.info("Reading taxonomy from taxtastic taxonomy and seqinfo files")
taxonomy_definition = gtns.read_taxtastic_taxonomy_and_seqinfo\
(open(taxtastic_taxonomy),
open(taxtastic_seqinfo))
else:
raise Exception("Taxonomy is required somehow e.g. by --taxonomy or --rerooted_annotated_tree")
# Check for duplicates
logging.info("Checking for duplicate sequences")
dup = self._check_for_duplicate_sequence_names(sequences)
if dup:
raise Exception("Found duplicate sequence name '%s' in sequences input file" % dup)
output_alignment_fh = tempfile.NamedTemporaryFile(prefix='graftm', suffix='.aln.faa')
tempfiles_to_close.append(output_alignment_fh)
output_alignment = output_alignment_fh.name
if user_hmm:
align_hmm = user_hmm
else:
align_hmm_fh = tempfile.NamedTemporaryFile(prefix='graftm', suffix='_align.hmm')
tempfiles_to_close.append(align_hmm_fh)
align_hmm = align_hmm_fh.name
if alignment:
dup = self._check_for_duplicate_sequence_names(alignment)
if dup:
raise Exception("Found duplicate sequence name '%s' in alignment input file" % dup)
ptype = self._get_hmm_from_alignment(alignment,
align_hmm,
output_alignment)
else:
logging.info("Aligning sequences to create aligned FASTA file")
ptype, output_alignment = self._align_and_create_hmm(sequences, alignment, user_hmm,
align_hmm, output_alignment, threads)
logging.info("Checking for incorrect or fragmented reads")
insufficiently_aligned_sequences = self._check_reads_hit(open(output_alignment),
min_aligned_percent)
while len(insufficiently_aligned_sequences) > 0:
logging.warn("One or more alignments do not span > %.2f %% of HMM" % (min_aligned_percent*100))
for s in insufficiently_aligned_sequences:
logging.warn("Insufficient alignment of %s, not including this sequence" % s)
sequences2_fh = tempfile.NamedTemporaryFile(prefix='graftm', suffix='.faa')
tempfiles_to_close.append(sequences2_fh)
sequences2 = sequences2_fh.name
num_sequences = self._remove_sequences_from_alignment(insufficiently_aligned_sequences,
sequences,
sequences2)
sequences = sequences2
if alignment:
alignment2_fh = tempfile.NamedTemporaryFile(prefix='graftm', suffix='.aln.faa')
tempfiles_to_close.append(alignment2_fh)
alignment2 = alignment2_fh.name
num_sequences = self._remove_sequences_from_alignment(insufficiently_aligned_sequences,
alignment,
alignment2)
alignment = alignment2
for name in insufficiently_aligned_sequences:
if rerooted_tree or rerooted_annotated_tree:
logging.warning('''Sequence %s in provided alignment does not meet the --min_aligned_percent cutoff. This sequence will be removed from the tree
in the final GraftM package. If you are sure these sequences are correct, turn off the --min_aligned_percent cutoff, provide it with a 0 (e.g. --min_aligned_percent 0) ''' % name)
removed_sequence_names.append(name)
logging.info("After removing %i insufficiently aligned sequences, left with %i sequences" % (len(insufficiently_aligned_sequences), num_sequences))
if num_sequences < 4:
raise Exception("Too few sequences remaining in alignment after removing insufficiently aligned sequences: %i" % num_sequences)
else:
logging.info("Reconstructing the alignment and HMM from remaining sequences")
output_alignment_fh = tempfile.NamedTemporaryFile(prefix='graftm', suffix='.aln.faa')
tempfiles_to_close.append(output_alignment_fh)
output_alignment = output_alignment_fh.name
if not user_hmm:
align_hmm_fh = tempfile.NamedTemporaryFile(prefix='graftm', suffix='.hmm')
tempfiles_to_close.append(align_hmm_fh)
align_hmm = align_hmm_fh.name
ptype, output_alignment= self._align_and_create_hmm(sequences, alignment, user_hmm,
align_hmm, output_alignment, threads)
logging.info("Checking for incorrect or fragmented reads")
insufficiently_aligned_sequences = self._check_reads_hit(open(output_alignment),
min_aligned_percent)
if not search_hmm_files:
search_hmm_fh = tempfile.NamedTemporaryFile(prefix='graftm', suffix='_search.hmm')
tempfiles_to_close.append(search_hmm_fh)
search_hmm = search_hmm_fh.name
self._create_search_hmm(sequences, taxonomy_definition, search_hmm, dereplication_level, threads)
search_hmm_files = [search_hmm]
# Make sure each sequence has been assigned a taxonomy:
aligned_sequence_objects = seqio.read_fasta_file(output_alignment)
unannotated = []
for s in aligned_sequence_objects:
if s.name not in taxonomy_definition:
unannotated.append(s.name)
if len(unannotated) > 0:
for s in unannotated:
logging.error("Unable to find sequence '%s' in the taxonomy definition" % s)
raise Exception("All sequences must be assigned a taxonomy, cannot continue")
logging.debug("Looking for non-standard characters in aligned sequences")
self._mask_strange_sequence_letters(aligned_sequence_objects, ptype)
# Deduplicate sequences - pplacer cannot handle these
logging.info("Deduplicating sequences")
dedup = Deduplicator()
deduplicated_arrays = dedup.deduplicate(aligned_sequence_objects)
deduplicated_taxonomy = dedup.lca_taxonomy(deduplicated_arrays, taxonomy_definition)
deduplicated_taxonomy_hash = {}
for i, tax in enumerate(deduplicated_taxonomy):
deduplicated_taxonomy_hash[deduplicated_arrays[i][0].name] = tax
deduplicated_alignment_file = base+"_deduplicated_aligned.fasta"
seqio.write_fasta_file([seqs[0] for seqs in deduplicated_arrays],
deduplicated_alignment_file)
logging.info("Removed %i sequences as duplicates, leaving %i non-identical sequences"\
% ((len(aligned_sequence_objects)-len(deduplicated_arrays)),
len(deduplicated_arrays)))
# Get corresponding unaligned sequences
filtered_names=[]
for list in [x for x in [x[1:] for x in deduplicated_arrays] if x]:
for seq in list:
filtered_names.append(seq.name)
sequences2_fh = tempfile.NamedTemporaryFile(prefix='graftm', suffix='.faa')
tempfiles_to_close.append(sequences2_fh)
sequences2 = sequences2_fh.name
# Create tree unless one was provided
if not rerooted_tree and not rerooted_annotated_tree and not unrooted_tree:
logging.debug("No tree provided")
logging.info("Building tree")
log_file, tre_file = self._build_tree(deduplicated_alignment_file,
base, ptype,
self.fasttree)
no_reroot = False
else:
if rerooted_tree:
logging.debug("Found unannotated pre-rerooted tree file %s" % rerooted_tree)
tre_file=rerooted_tree
no_reroot = True
elif rerooted_annotated_tree:
logging.debug("Found annotated pre-rerooted tree file %s" % rerooted_tree)
tre_file=rerooted_annotated_tree
no_reroot = True
elif unrooted_tree:
logging.info("Using input unrooted tree")
tre_file = unrooted_tree
no_reroot = False
else:
raise
# Remove any sequences from the tree that are duplicates
cleaner = DendropyTreeCleaner()
tree = Tree.get(path=tre_file, schema='newick')
for group in deduplicated_arrays:
[removed_sequence_names.append(s.name) for s in group[1:]]
cleaner.remove_sequences(tree, removed_sequence_names)
# Ensure there is nothing amiss now as a user-interface thing
cleaner.match_alignment_and_tree_sequence_ids(\
[g[0].name for g in deduplicated_arrays], tree)
if tree_log:
# User specified a log file, go with that
logging.debug("Using user-specified log file %s" % tree_log)
log_file = tree_log
else:
logging.info("Generating log file")
log_file_tempfile = tempfile.NamedTemporaryFile(suffix='.tree_log', prefix='graftm')
tempfiles_to_close.append(log_file_tempfile)
log_file = log_file_tempfile.name
tre_file_tempfile = tempfile.NamedTemporaryFile(suffix='.tree', prefix='graftm')
tempfiles_to_close.append(tre_file_tempfile)
tre_file = tre_file_tempfile.name
with tempfile.NamedTemporaryFile(suffix='.tree', prefix='graftm') as f:
# Make the newick file simple (ie. un-arb it) for fasttree.
cleaner.write_fasttree_newick(tree, f)
f.flush()
self._generate_tree_log_file(f.name, deduplicated_alignment_file,
tre_file, log_file, ptype, self.fasttree)
# Create tax and seqinfo .csv files
taxonomy_to_keep=[
seq.name for seq in
[x for x in [x[0] for x in deduplicated_arrays]
if x]
]
refpkg = "%s.refpkg" % output_gpkg_path
self.the_trash.append(refpkg)
if taxtastic_taxonomy and taxtastic_seqinfo:
logging.info("Creating reference package")
refpkg = self._taxit_create(base, deduplicated_alignment_file,
tre_file, log_file, taxtastic_taxonomy,
taxtastic_seqinfo, refpkg, no_reroot)
else:
gtns = Getaxnseq()
seq = base+"_seqinfo.csv"
tax = base+"_taxonomy.csv"
self.the_trash += [seq, tax]
if rerooted_annotated_tree:
logging.info("Building seqinfo and taxonomy file from input annotated tree")
taxonomy_definition = TaxonomyExtractor().taxonomy_from_annotated_tree(
Tree.get(path=rerooted_annotated_tree, schema='newick'))
elif taxonomy:
logging.info("Building seqinfo and taxonomy file from input taxonomy")
taxonomy_definition = GreenGenesTaxonomy.read_file(taxonomy).taxonomy
else:
raise Exception("Programming error: Taxonomy is required somehow e.g. by --taxonomy or --rerooted_annotated_tree")
taxonomy_definition = {x:taxonomy_definition[x]
for x in taxonomy_definition
if x in taxonomy_to_keep}
gtns.write_taxonomy_and_seqinfo_files(taxonomy_definition,
tax,
seq)
# Create the reference package
logging.info("Creating reference package")
refpkg = self._taxit_create(base, deduplicated_alignment_file,
tre_file, log_file, tax, seq, refpkg,
no_reroot)
if sequences:
# Run diamond makedb
logging.info("Creating diamond database")
if ptype == Create._PROTEIN_PACKAGE_TYPE:
cmd = "diamond makedb --in '%s' -d '%s'" % (sequences, base)
extern.run(cmd)
diamondb = '%s.dmnd' % base
elif ptype == Create._NUCLEOTIDE_PACKAGE_TYPE:
diamondb = None
else: raise Exception("Programming error")
else:
diamondb = None
if sequences:
# Get range
max_range = self._define_range(sequences)
else:
max_range = self._define_range(alignment)
# Compile the gpkg
logging.info("Compiling gpkg")
GraftMPackageVersion3.compile(output_gpkg_path, refpkg, align_hmm, diamondb,
max_range, sequences, search_hmm_files=search_hmm_files)
logging.info("Cleaning up")
self._cleanup(self.the_trash)
for tf in tempfiles_to_close:
tf.close()
# Test out the gpkg just to be sure.
#
# TODO: Use graftM through internal means rather than via extern. This
# requires some refactoring so that graft() can be called easily with
# sane defaults.
logging.info("Testing gpkg package works")
self._test_package(output_gpkg_path)
logging.info("Finished\n")
def main(self, **kwargs):
alignment = kwargs.pop('alignment', None)
sequences = kwargs.pop('sequences', None)
taxonomy = kwargs.pop('taxonomy', None)
rerooted_tree = kwargs.pop('rerooted_tree', None)
unrooted_tree = kwargs.pop('unrooted_tree', None)
tree_log = kwargs.pop('tree_log', None)
prefix = kwargs.pop('prefix', None)
rerooted_annotated_tree = kwargs.pop('rerooted_annotated_tree', None)
user_hmm = kwargs.pop('hmm', None)
search_hmm_files = kwargs.pop('search_hmm_files', None)
min_aligned_percent = kwargs.pop('min_aligned_percent', 0.01)
taxtastic_taxonomy = kwargs.pop('taxtastic_taxonomy', None)
taxtastic_seqinfo = kwargs.pop('taxtastic_seqinfo', None)
force_overwrite = kwargs.pop('force', False)
graftm_package = kwargs.pop('graftm_package', False)
dereplication_level = kwargs.pop('dereplication_level', False)
threads = kwargs.pop('threads', 5)
if len(kwargs) > 0:
raise Exception("Unexpected arguments detected: %s" % kwargs)
seqio = SequenceIO()
locus_name = (os.path.basename(sequences).split('.')[0] if sequences
else os.path.basename(alignment).split('.')[0])
tmp = tempdir.TempDir()
base = os.path.join(tmp.name, locus_name)
insufficiently_aligned_sequences = [None]
removed_sequence_names = []
if prefix:
output_gpkg_path = prefix
else:
output_gpkg_path = "%s.gpkg" % locus_name
if os.path.exists(output_gpkg_path):
if force_overwrite:
logging.warn("Deleting previous directory %s" %
output_gpkg_path)
shutil.rmtree(output_gpkg_path)
else:
raise Exception(
"Cowardly refusing to overwrite gpkg to already existing %s"
% output_gpkg_path)
logging.info("Building gpkg for %s" % output_gpkg_path)
# Read in taxonomy somehow
gtns = Getaxnseq()
if rerooted_annotated_tree:
logging.info(
"Building seqinfo and taxonomy file from input annotated tree")
taxonomy_definition = TaxonomyExtractor().taxonomy_from_annotated_tree(\
Tree.get(path=rerooted_annotated_tree, schema='newick'))
elif taxonomy:
logging.info(
"Building seqinfo and taxonomy file from input taxonomy")
taxonomy_definition = GreenGenesTaxonomy.read_file(
taxonomy).taxonomy
elif taxtastic_seqinfo and taxtastic_taxonomy:
logging.info(
"Reading taxonomy from taxtastic taxonomy and seqinfo files")
taxonomy_definition = gtns.read_taxtastic_taxonomy_and_seqinfo\
(open(taxtastic_taxonomy),
open(taxtastic_seqinfo))
else:
raise Exception(
"Taxonomy is required somehow e.g. by --taxonomy or --rerooted_annotated_tree"
)
# Check for duplicates
logging.info("Checking for duplicate sequences")
dup = self._check_for_duplicate_sequence_names(sequences)
if dup:
raise Exception(
"Found duplicate sequence name '%s' in sequences input file" %
dup)
output_alignment = tempfile.NamedTemporaryFile(prefix='graftm',
suffix='.aln.faa').name
align_hmm = (user_hmm if user_hmm else tempfile.NamedTemporaryFile(
prefix='graftm', suffix='_align.hmm').name)
if alignment:
dup = self._check_for_duplicate_sequence_names(alignment)
if dup:
raise Exception(
"Found duplicate sequence name '%s' in alignment input file"
% dup)
ptype = self._get_hmm_from_alignment(alignment, align_hmm,
output_alignment)
else:
logging.info("Aligning sequences to create aligned FASTA file")
ptype, output_alignment = self._align_and_create_hmm(
sequences, alignment, user_hmm, align_hmm, output_alignment,
threads)
logging.info("Checking for incorrect or fragmented reads")
insufficiently_aligned_sequences = self._check_reads_hit(
open(output_alignment), min_aligned_percent)
while len(insufficiently_aligned_sequences) > 0:
logging.warn(
"One or more alignments do not span > %.2f %% of HMM" %
(min_aligned_percent * 100))
for s in insufficiently_aligned_sequences:
logging.warn(
"Insufficient alignment of %s, not including this sequence"
% s)
_, sequences2 = tempfile.mkstemp(prefix='graftm', suffix='.faa')
num_sequences = self._remove_sequences_from_alignment(
insufficiently_aligned_sequences, sequences, sequences2)
sequences = sequences2
if alignment:
_, alignment2 = tempfile.mkstemp(prefix='graftm',
suffix='.aln.faa')
num_sequences = self._remove_sequences_from_alignment(
insufficiently_aligned_sequences, alignment, alignment2)
alignment = alignment2
for name in insufficiently_aligned_sequences:
if rerooted_tree or rerooted_annotated_tree:
logging.warning(
'''Sequence %s in provided alignment does not meet the --min_aligned_percent cutoff. This sequence will be removed from the tree
in the final GraftM package. If you are sure these sequences are correct, turn off the --min_aligned_percent cutoff, provide it with a 0 (e.g. --min_aligned_percent 0) '''
% name)
removed_sequence_names.append(name)
logging.info(
"After removing %i insufficiently aligned sequences, left with %i sequences"
% (len(insufficiently_aligned_sequences), num_sequences))
if num_sequences < 4:
raise Exception(
"Too few sequences remaining in alignment after removing insufficiently aligned sequences: %i"
% num_sequences)
else:
logging.info(
"Reconstructing the alignment and HMM from remaining sequences"
)
output_alignment = tempfile.NamedTemporaryFile(
prefix='graftm', suffix='.aln.faa').name
if not user_hmm:
align_hmm = tempfile.NamedTemporaryFile(prefix='graftm',
suffix='.hmm').name
ptype, output_alignment = self._align_and_create_hmm(
sequences, alignment, user_hmm, align_hmm,
output_alignment, threads)
logging.info("Checking for incorrect or fragmented reads")
insufficiently_aligned_sequences = self._check_reads_hit(
open(output_alignment), min_aligned_percent)
if not search_hmm_files:
search_hmm = tempfile.NamedTemporaryFile(prefix='graftm',
suffix='_search.hmm').name
self._create_search_hmm(sequences, taxonomy_definition, search_hmm,
dereplication_level, threads)
search_hmm_files = [search_hmm]
# Make sure each sequence has been assigned a taxonomy:
aligned_sequence_objects = seqio.read_fasta_file(output_alignment)
unannotated = []
for s in aligned_sequence_objects:
if s.name not in taxonomy_definition:
unannotated.append(s.name)
if len(unannotated) > 0:
for s in unannotated:
logging.error(
"Unable to find sequence '%s' in the taxonomy definition" %
s)
raise Exception(
"All sequences must be assigned a taxonomy, cannot continue")
logging.debug(
"Looking for non-standard characters in aligned sequences")
self._mask_strange_sequence_letters(aligned_sequence_objects, ptype)
# Deduplicate sequences - pplacer cannot handle these
logging.info("Deduplicating sequences")
dedup = Deduplicator()
deduplicated_arrays = dedup.deduplicate(aligned_sequence_objects)
deduplicated_taxonomy = dedup.lca_taxonomy(deduplicated_arrays,
taxonomy_definition)
deduplicated_taxonomy_hash = {}
for i, tax in enumerate(deduplicated_taxonomy):
deduplicated_taxonomy_hash[deduplicated_arrays[i][0].name] = tax
deduplicated_alignment_file = base + "_deduplicated_aligned.fasta"
seqio.write_fasta_file([seqs[0] for seqs in deduplicated_arrays],
deduplicated_alignment_file)
logging.info("Removed %i sequences as duplicates, leaving %i non-identical sequences"\
% ((len(aligned_sequence_objects)-len(deduplicated_arrays)),
len(deduplicated_arrays)))
# Get corresponding unaligned sequences
filtered_names = []
for list in [x for x in [x[1:] for x in deduplicated_arrays] if x]:
for seq in list:
filtered_names.append(seq.name)
_, sequences2 = tempfile.mkstemp(prefix='graftm', suffix='.faa')
# Create tree unless one was provided
if not rerooted_tree and not rerooted_annotated_tree and not unrooted_tree:
logging.debug("No tree provided")
logging.info("Building tree")
log_file, tre_file = self._build_tree(deduplicated_alignment_file,
base, ptype, self.fasttree)
no_reroot = False
else:
if rerooted_tree:
logging.debug("Found unannotated pre-rerooted tree file %s" %
rerooted_tree)
tre_file = rerooted_tree
no_reroot = True
elif rerooted_annotated_tree:
logging.debug("Found annotated pre-rerooted tree file %s" %
rerooted_tree)
tre_file = rerooted_annotated_tree
no_reroot = True
elif unrooted_tree:
logging.info("Using input unrooted tree")
tre_file = unrooted_tree
no_reroot = False
else:
raise
# Remove any sequences from the tree that are duplicates
cleaner = DendropyTreeCleaner()
tree = Tree.get(path=tre_file, schema='newick')
for group in deduplicated_arrays:
[removed_sequence_names.append(s.name) for s in group[1:]]
cleaner.remove_sequences(tree, removed_sequence_names)
# Ensure there is nothing amiss now as a user-interface thing
cleaner.match_alignment_and_tree_sequence_ids(\
[g[0].name for g in deduplicated_arrays], tree)
if tree_log:
# User specified a log file, go with that
logging.debug("Using user-specified log file %s" % tree_log)
log_file = tree_log
else:
logging.info("Generating log file")
log_file_tempfile = tempfile.NamedTemporaryFile(
suffix='.tree_log', prefix='graftm')
log_file = log_file_tempfile.name
tre_file_tempfile = tempfile.NamedTemporaryFile(
suffix='.tree', prefix='graftm')
tre_file = tre_file_tempfile.name
with tempfile.NamedTemporaryFile(suffix='.tree',
prefix='graftm') as f:
# Make the newick file simple (ie. un-arb it) for fasttree.
cleaner.write_fasttree_newick(tree, f)
f.flush()
self._generate_tree_log_file(f.name,
deduplicated_alignment_file,
tre_file, log_file, ptype,
self.fasttree)
# Create tax and seqinfo .csv files
taxonomy_to_keep = [
seq.name
for seq in [x for x in [x[0] for x in deduplicated_arrays] if x]
]
refpkg = "%s.refpkg" % output_gpkg_path
self.the_trash.append(refpkg)
if taxtastic_taxonomy and taxtastic_seqinfo:
logging.info("Creating reference package")
refpkg = self._taxit_create(base, deduplicated_alignment_file,
tre_file, log_file, taxtastic_taxonomy,
taxtastic_seqinfo, refpkg, no_reroot)
else:
gtns = Getaxnseq()
seq = base + "_seqinfo.csv"
tax = base + "_taxonomy.csv"
self.the_trash += [seq, tax]
if rerooted_annotated_tree:
logging.info(
"Building seqinfo and taxonomy file from input annotated tree"
)
taxonomy_definition = TaxonomyExtractor(
).taxonomy_from_annotated_tree(
Tree.get(path=rerooted_annotated_tree, schema='newick'))
elif taxonomy:
logging.info(
"Building seqinfo and taxonomy file from input taxonomy")
taxonomy_definition = GreenGenesTaxonomy.read_file(
taxonomy).taxonomy
else:
raise Exception(
"Programming error: Taxonomy is required somehow e.g. by --taxonomy or --rerooted_annotated_tree"
)
taxonomy_definition = {
x: taxonomy_definition[x]
for x in taxonomy_definition if x in taxonomy_to_keep
}
gtns.write_taxonomy_and_seqinfo_files(taxonomy_definition, tax,
seq)
# Create the reference package
logging.info("Creating reference package")
refpkg = self._taxit_create(base, deduplicated_alignment_file,
tre_file, log_file, tax, seq, refpkg,
no_reroot)
if sequences:
# Run diamond makedb
logging.info("Creating diamond database")
if ptype == Create._PROTEIN_PACKAGE_TYPE:
cmd = "diamond makedb --in '%s' -d '%s'" % (sequences, base)
extern.run(cmd)
diamondb = '%s.dmnd' % base
elif ptype == Create._NUCLEOTIDE_PACKAGE_TYPE:
diamondb = None
else:
raise Exception("Programming error")
else:
diamondb = None
if sequences:
# Get range
max_range = self._define_range(sequences)
else:
max_range = self._define_range(alignment)
# Compile the gpkg
logging.info("Compiling gpkg")
GraftMPackageVersion3.compile(output_gpkg_path,
refpkg,
align_hmm,
diamondb,
max_range,
sequences,
search_hmm_files=search_hmm_files)
logging.info("Cleaning up")
self._cleanup(self.the_trash)
# Test out the gpkg just to be sure.
#
# TODO: Use graftM through internal means rather than via extern. This
# requires some refactoring so that graft() can be called easily with
# sane defaults.
logging.info("Testing gpkg package works")
self._test_package(output_gpkg_path)
logging.info("Finished\n")
def _assign_taxonomy_with_diamond(self, base_list, db_search_results,
graftm_package, graftm_files,
diamond_performance_parameters):
'''Run diamond to assign taxonomy
Parameters
----------
base_list: list of str
list of sequence block names
db_search_results: list of DBSearchResult
the result of running hmmsearches
graftm_package: GraftMPackage object
Diamond is run against this database
graftm_files: GraftMFiles object
Result files are written here
diamond_performance_parameters : str
extra args for DIAMOND
Returns
-------
list of
1. time taken for assignment
2. assignments i.e. dict of base_list entry to dict of read names to
to taxonomies, or None if there was no hit detected.
'''
runner = Diamond(graftm_package.diamond_database_path(),
self.args.threads, self.args.evalue)
taxonomy_definition = Getaxnseq().read_taxtastic_taxonomy_and_seqinfo\
(open(graftm_package.taxtastic_taxonomy_path()),
open(graftm_package.taxtastic_seqinfo_path()))
results = {}
# For each of the search results,
for i, search_result in enumerate(db_search_results):
if search_result.hit_fasta() is None:
sequence_id_to_taxonomy = {}
else:
sequence_id_to_hit = {}
# Run diamond
logging.debug("Running diamond on %s" %
search_result.hit_fasta())
diamond_result = runner.run(
search_result.hit_fasta(),
UnpackRawReads.PROTEIN_SEQUENCE_TYPE,
daa_file_basename=graftm_files.
diamond_assignment_output_basename(base_list[i]),
extra_args=diamond_performance_parameters)
for res in diamond_result.each([
SequenceSearchResult.QUERY_ID_FIELD,
SequenceSearchResult.HIT_ID_FIELD
]):
if res[0] in sequence_id_to_hit:
# do not accept duplicates
if sequence_id_to_hit[res[0]] != res[1]:
raise Exception(
"Diamond unexpectedly gave two hits for a single query sequence for %s"
% res[0])
else:
sequence_id_to_hit[res[0]] = res[1]
# Extract taxonomy of the best hit, and add in the no hits
sequence_id_to_taxonomy = {}
for seqio in SequenceIO().read_fasta_file(
search_result.hit_fasta()):
name = seqio.name
if name in sequence_id_to_hit:
# Add Root; to be in line with pplacer assignment method
sequence_id_to_taxonomy[name] = [
'Root'
] + taxonomy_definition[sequence_id_to_hit[name]]
else:
# picked up in the initial search (by hmmsearch, say), but diamond misses it
sequence_id_to_taxonomy[name] = ['Root']
results[base_list[i]] = sequence_id_to_taxonomy
return results