| name | bio-comparative-genomics-positive-selection |
| description | Detect positive selection using dN/dS (omega) tests with PAML codeml and HyPhy. Identify sites and branches under adaptive evolution through codon models and branch-site tests. Use when testing for adaptive evolution in gene families or identifying positively selected sites. |
| tool_type | mixed |
| primary_tool | PAML |
Positive Selection Analysis
dN/dS Overview
'''
dN/dS (omega, ω) interpretation:
- ω < 1: Purifying (negative) selection - deleterious mutations removed
- ω = 1: Neutral evolution - no selective pressure
- ω > 1: Positive (diversifying) selection - advantageous mutations favored
Most genes: ω << 1 (strong purifying selection)
Immune genes, reproduction: Often show ω > 1 at specific sites
'''
PAML Codeml Analysis
'''Run PAML codeml for selection analysis'''
import subprocess
import os
from Bio import SeqIO
from Bio.Seq import Seq
def prepare_codon_alignment(cds_fasta, output_phy):
'''Prepare codon alignment in PHYLIP format
Requirements:
- CDS sequences (in-frame, no stop codons except terminal)
- Multiple sequence alignment already performed
- Sequence length divisible by 3
'''
records = list(SeqIO.parse(cds_fasta, 'fasta'))
for rec in records:
if len(rec.seq) % 3 != 0:
print(f'Warning: {rec.id} length not divisible by 3')
n_seq = len(records)
seq_len = len(records[0].seq)
with open(output_phy, 'w') as f:
f.write(f' {n_seq} {seq_len}\n')
for rec in records:
name = rec.id[:10].ljust(10)
f.write(f'{name}{str(rec.seq)}\n')
return output_phy
def create_codeml_control(alignment_file, tree_file, output_dir, model='M8'):
'''Create codeml control file
Site models for detecting positive selection:
- M0: One ratio (single ω for all sites)
- M1a: Nearly neutral (ω0 < 1, ω1 = 1)
- M2a: Positive selection (ω0 < 1, ω1 = 1, ω2 > 1)
- M7: Beta (ω from beta distribution, 0 < ω < 1)
- M8: Beta + ω > 1 (allows positive selection)
- M8a: Beta + ω = 1 (null for M8 comparison)
Standard comparison: M8 vs M7 or M8 vs M8a
'''
model_params = {
'M0': {'NSsites': 0, 'model': 0},
'M1a': {'NSsites': 1, 'model': 0},
'M2a': {'NSsites': 2, 'model': 0},
'M7': {'NSsites': 7, 'model': 0},
'M8': {'NSsites': 8, 'model': 0},
'M8a': {'NSsites': 8, 'model': 0, 'fix_omega': 1, 'omega': 1},
}
params = model_params.get(model, model_params['M8'])
ctl_content = f'''
seqfile = {alignment_file}
treefile = {tree_file}
outfile = {output_dir}/mlc
noisy = 9
verbose = 1
runmode = 0
seqtype = 1
CodonFreq = 2
model = {params.get('model', 0)}
NSsites = {params.get('NSsites', 8)}
icode = 0
fix_kappa = 0
kappa = 2
fix_omega = {params.get('fix_omega', 0)}
omega = {params.get('omega', 1)}
'''
ctl_file = f'{output_dir}/codeml_{model}.ctl'
with open(ctl_file, 'w') as f:
f.write(ctl_content)
return ctl_file
def run_codeml(ctl_file):
'''Run PAML codeml'''
cmd = f'codeml {ctl_file}'
result = subprocess.run(cmd, shell=True, capture_output=True, text=True)
if result.returncode != 0:
print(f'Codeml error: {result.stderr}')
return result
def parse_codeml_output(mlc_file):
'''Parse codeml output for likelihood and parameters'''
results = {'lnL': None, 'omega': None, 'kappa': None, 'sites': []}
with open(mlc_file) as f:
content = f.read()
for line in content.split('\n'):
if 'lnL' in line and 'np' in line:
parts = line.split()
for i, p in enumerate(parts):
if p == 'lnL':
results['lnL'] = float(parts[i + 2])
break
if 'omega' in line.lower() and '=' in line:
parts = line.split('=')
if len(parts) >= 2:
try:
results['omega'] = float(parts[-1].strip().split()[0])
except ValueError:
pass
if 'Bayes Empirical Bayes' in content:
beb_section = content.split('Bayes Empirical Bayes')[1]
for line in beb_section.split('\n'):
parts = line.split()
if len(parts) >= 5:
try:
site = int(parts[0])
aa = parts[1]
prob = float(parts[2])
if prob > 0.95:
results['sites'].append({
'position': site,
'amino_acid': aa,
'probability': prob,
'significance': '**' if prob > 0.99 else '*'
})
except (ValueError, IndexError):
continue
return results
def likelihood_ratio_test(lnL_null, lnL_alt, df=2):
'''Perform likelihood ratio test
For M8 vs M7: df = 2
For M2a vs M1a: df = 2
For branch-site test: df = 1
Significance thresholds (chi-square):
- df=1: 3.84 (p<0.05), 6.63 (p<0.01)
- df=2: 5.99 (p<0.05), 9.21 (p<0.01)
'''
from scipy import stats
lrt = 2 * (lnL_alt - lnL_null)
p_value = 1 - stats.chi2.cdf(lrt, df)
return {
'LRT_statistic': lrt,
'degrees_freedom': df,
'p_value': p_value,
'significant': p_value < 0.05
}
Branch-Site Models
def create_branch_site_control(alignment, tree, foreground_branch, output_dir):
'''Create control file for branch-site test
Branch-site model A tests for positive selection on
specific branches (foreground) while allowing variation
across the tree.
Foreground branch: Mark with #1 in tree file
Example: ((A,B),(C #1,D));
Comparison: Model A vs null (fix_omega = 1)
'''
ctl_content = f'''
seqfile = {alignment}
treefile = {tree}
outfile = {output_dir}/branch_site.mlc
runmode = 0
seqtype = 1
CodonFreq = 2
model = 2
NSsites = 2
fix_kappa = 0
kappa = 2
fix_omega = 0
omega = 1
'''
ctl_file = f'{output_dir}/branch_site.ctl'
with open(ctl_file, 'w') as f:
f.write(ctl_content)
return ctl_file
def mark_foreground_branch(tree_file, foreground_taxa, output_file):
'''Mark foreground lineage in Newick tree
For testing selection on specific lineage:
- Add #1 after the clade of interest
- Can mark multiple branches with different tags
'''
with open(tree_file) as f:
tree = f.read().strip()
for taxon in foreground_taxa:
tree = tree.replace(taxon, f'{taxon} #1')
with open(output_file, 'w') as f:
f.write(tree)
return output_file
HyPhy Alternative
def run_hyphy_busted(alignment_file, tree_file, output_json):
'''Run HyPhy BUSTED for gene-wide selection
BUSTED (Branch-site Unrestricted Statistical Test for
Episodic Diversification) tests whether a gene has
experienced positive selection at any site on any branch.
More sensitive than PAML for episodic selection
'''
cmd = f'hyphy busted --alignment {alignment_file} --tree {tree_file} --output {output_json}'
subprocess.run(cmd, shell=True)
return output_json
def run_hyphy_meme(alignment_file, tree_file, output_json):
'''Run HyPhy MEME for site-specific selection
MEME (Mixed Effects Model of Evolution) detects
episodic diversifying selection at individual sites.
Better than PAML M8 when selection is episodic
(not constant across all branches)
'''
cmd = f'hyphy meme --alignment {alignment_file} --tree {tree_file} --output {output_json}'
subprocess.run(cmd, shell=True)
return output_json
def run_hyphy_fel(alignment_file, tree_file, output_json):
'''Run HyPhy FEL for pervasive selection
FEL (Fixed Effects Likelihood) tests for pervasive
selection at each site across the entire phylogeny.
Use when selection is expected to be constant
'''
cmd = f'hyphy fel --alignment {alignment_file} --tree {tree_file} --output {output_json}'
subprocess.run(cmd, shell=True)
return output_json
def parse_hyphy_json(json_file):
'''Parse HyPhy JSON output'''
import json
(json_file) f:
results = json.load(f)
test_results = results.get(, {})
sites = []
mle = results.get(, {}).get(, {})
site_data mle.get(, {}).values():
(site_data, ) (site_data) > :
{
: test_results.get(),
: test_results.get(),
: sites
}
Related Skills
- comparative-genomics/synteny-analysis - Find syntenic genes for selection tests
- comparative-genomics/ortholog-inference - Identify orthologs for analysis
- alignment/msa-parsing - Parse and manipulate codon alignments
- phylogenetics/modern-tree-inference - Generate trees for codeml
