Class: BacterialComparator

Inherits:
Object
  • Object
show all
Defined in:
lib/bacterial-comparator.rb

Instance Attribute Summary collapse

Instance Method Summary collapse

Constructor Details

#initialize(options, root) ⇒ BacterialComparator

Initialize BacterialAnnotator options, options, ROOT, options, options)



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# File 'lib/bacterial-comparator.rb', line 20

def initialize options, root

  @root = root
  @outdir = options[:outdir]
  Dir.mkdir(@outdir) if ! Dir.exists? @outdir
  @genomes_list = options[:genomes_list]
  @proc = options[:proc].to_i
  @phylo_nb_genes = options[:phylo_nb_genes]
  min_cov = options[:min_cov].to_f
  min_pid = options[:pidentity].to_f
  if min_cov > 1
    min_cov = min_cov/100
  end
  if min_pid > 1
    min_pid = min_pid/100
  end

  @ref_prot = get_ref_prot
  @synteny = read_prot_synteny
  @stats = extract_syntenic_fasta min_cov, min_pid

end

Instance Attribute Details

#genomes_list ⇒ Object (readonly)

Returns the value of attribute genomes_list.



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# File 'lib/bacterial-comparator.rb', line 16

def genomes_list
  @genomes_list
end

#stats ⇒ Object (readonly)

Returns the value of attribute stats.



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# File 'lib/bacterial-comparator.rb', line 16

def stats
  @stats
end

Instance Method Details

#build_multifasta(synteny_list) ⇒ Object 



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# File 'lib/bacterial-comparator.rb', line 103

def build_multifasta synteny_list

  pep_out_dir = "./#{@outdir}/align-genes-pep"

  ref_proteins = load_genome_cds(Dir["#{@genomes_list[0]}/*.pep"][0])
  synteny_list.each do |k,v|
    pep_out = File.open(pep_out_dir+"/#{k}.pep", "w")
    pep_out.write(ref_proteins[k])
    pep_out.close
  end

  @genomes_list.each_with_index do |g,i|

    genome_proteins = load_genome_cds("#{g}/Proteins.fa")
    synteny_list.each do |k,v|
      pep_out = File.open(pep_out_dir+"/#{k}.pep", "a")
      pep_out.write(genome_proteins[v[i][:query_prot]])
      pep_out.close
    end

  end

  dna_out_dir = "./#{@outdir}/align-genes-dna"
  ref_genes = load_genome_cds(Dir["#{@genomes_list[0]}/*.dna"][0])
  synteny_list.each do |k,v|
    dna_out = File.open(dna_out_dir+"/#{k}.dna", "w")
    dna_out.write(ref_genes[k])
    dna_out.close
  end

  @genomes_list.each_with_index do |g,i|

    genome_genes = load_genome_cds("#{g}/Genes.fa")
    synteny_list.each do |k,v|
      dna_out = File.open(dna_out_dir+"/#{k}.dna", "a")
      dna_out.write(genome_genes[v[i][:query_prot]])
      dna_out.close
    end

  end

end

#concat_alignments(outfile, ref_id) ⇒ Object 



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# File 'lib/bacterial-comparator.rb', line 326

def concat_alignments outfile, ref_id

  if File.exists?("../#{outfile}") and File.size("../#{outfile}") > 0
    puts "..Alignment concatenated file already exists, skipping."
    return
  end

  fout = File.open("../#{outfile}", "w")

  seq = ""
  Dir["*.aln"].each do |f|
    flat = Bio::FlatFile.auto(f)
    ref_seq = flat.entries[0]
    flat.close
    seq += ref_seq.seq
  end

  bioseq = Bio::Sequence.auto(seq)
  out = bioseq.output_fasta("#{ref_id}", 60)
  fout.write(out)

  for i in 1..@genomes_list.length
    seq = ""
    Dir["*.aln"].each do |f|
      flat = Bio::FlatFile.auto(f)
      j=0
      flat.each_entry do |entry|
        if j<i
          j+=1
          next
        elsif i == j
          seq += entry.seq
          j+=1
        else
          break
        end
      end
      flat.close
    end
    bioseq = Bio::Sequence.auto(seq)
    # get the file name without path prefix and extension
    genome_name = genomes_list[i-1].split("/")[-1].split(".")[0]
    out = bioseq.output_fasta(genome_name,60)
    fout.write(out)
  end

  fout.close

end

#extract_syntenic_fasta(min_cov, min_pid) ⇒ Object

extract and dump multifasta for syntenic genes and proteins



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# File 'lib/bacterial-comparator.rb', line 147

def extract_syntenic_fasta min_cov, min_pid

  print "# Extracting Proteins and Genes multifasta.."
  start_time = Time.now

  nb_of_syntenic = 0
  stats = {}
  stats[:syntenic] = []
  fout = File.open("#{@outdir}/cds-synteny.tsv", "w")
  fout.write("Gene\t"+@genomes_list.join("\t")+"\n")

  to_build_multifasta = []

  @synteny.each do |k,v|

    is_syntenic = 1
    v.each do |v_|
      if v_[:query_cov] == "-"
        is_syntenic = 0
        break
      elsif v_[:query_cov] > min_cov and
           v_[:ref_cov] > min_cov and
           v_[:pId] > min_pid
      # synteny -> great !
      else
        is_syntenic = 0
        break
      end
    end

    fout.write("#{k}")
    if is_syntenic == 1
      nb_of_syntenic += 1
      # build_multifasta k, v
      to_build_multifasta << [k,v]
      v.each do |x|
        fout.write("\t#{x[:query_prot]}|#{x[:pId]}|#{x[:query_cov]}|#{x[:ref_cov]}")
        stats[:syntenic] << k
      end
      fout.write("\n")
    else
      v.each do |x|
        if x[:pId] == 0.0
          fout.write("\t-")
        else
          fout.write("\t#{x[:query_prot]}|#{x[:pId]}|#{x[:query_cov]}|#{x[:ref_cov]}")
        end
      end
      fout.write("\n")
    end

  end

  fout.close

  pep_out_dir = "./#{@outdir}/align-genes-pep"
  dna_out_dir = "./#{@outdir}/align-genes-dna"
  Dir.mkdir(pep_out_dir) if ! Dir.exists? pep_out_dir
  Dir.mkdir(dna_out_dir) if ! Dir.exists? dna_out_dir

  synteny_list = to_build_multifasta.each_slice((to_build_multifasta.length/@proc)+1).to_a

  Parallel.map(synteny_list, in_processes: @proc) { |list|
    build_multifasta list
  }

  end_time = Time.now
  c_time = Helper.sec2str(end_time-start_time)
  print "done (#{c_time})\n"

  stats[:nb_of_syntenic] = nb_of_syntenic
  #puts "   Syntenic genes : " + nb_of_syntenic.to_s + " / " + @ref_prot.length.to_s

end

#get_ref_prot ⇒ Object 



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# File 'lib/bacterial-comparator.rb', line 74

def get_ref_prot
  ref_prot = []
  pep_file = Dir["#{@genomes_list[0]}/*.pep"]
  flatfile = Bio::FlatFile.auto("#{pep_file[0]}")
  flatfile.each_entry do |entry|
    ref_prot << entry.definition.split(" ")[0]
  end
  flatfile.close
  ref_prot
end

#load_genome_cds(file) ⇒ Object

load all id => sequences from multifasta



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# File 'lib/bacterial-comparator.rb', line 87

def load_genome_cds file

  proteins = {}
  flatfile = Bio::FlatFile.auto(file)
  flatfile.each_entry do |entry|
    name = entry.definition.split(" ")[0]
    bioseq = Bio::Sequence.auto(entry.seq)
    out = bioseq.output_fasta("#{name}",60)
    proteins[name] = out
  end

  proteins

end

#mafft_align(f) ⇒ Object 



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# File 'lib/bacterial-comparator.rb', line 223

def mafft_align f

  trying = 0
  begin
    cmd = system("#{@root}/mafft.linux --quiet #{f} > #{f}.aln")
    if File.size("#{f}.aln") == 0
      puts "File size of 0.. --#{f}--"
      puts "Command used : #{@root}/mafft.linux --quiet #{f} > #{f}.aln"
      fail
    else
      status = "OK"
      status = "FAILED" if cmd != true
      # puts "Alignment #{f} : #{status}"
    end
  rescue
    if trying < 3
      trying += 1
      retry
    end
    status = "FAILED"
    puts "Alignment #{f} : #{status}"
  end

end

#mafft_align_all_dna ⇒ Object 



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# File 'lib/bacterial-comparator.rb', line 287

def mafft_align_all_dna

  print "# Sequence alignments - conserved single genes dna (MAFFT).."
  start_time = Time.now

  ori_dir = Dir.pwd
  Dir.chdir("#{@outdir}/align-genes-dna/")

  is_done = 1
  if Dir["*.dna"].length == Dir["*.aln"].length
    Dir["*.aln"].each do |a|
      if File.size(a) == 0
        is_done = 0
      end
    end
  else
    is_done = 0
  end

  if is_done == 0
    Parallel.map(Dir["*.dna"], in_processes: @proc) { |f|
      mafft_align f
    }
  end

  # ugly hack to find out the reference genome
  ref_id = Dir["#{ori_dir}/#{@genomes_list[0]}/*.pep"][0].split('/')[-1].gsub(".pep","")

  end_time = Time.now
  c_time = Helper.sec2str(end_time-start_time)
  print "done (#{c_time})\n"

  concat_alignments "align-genes-dna.all.fasta", ref_id

  Dir.chdir(ori_dir)

end

#mafft_align_all_pep ⇒ Object 



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# File 'lib/bacterial-comparator.rb', line 249

def mafft_align_all_pep

  print "# Sequence alignments - conserved single proteins a.a. (MAFFT).."
  start_time = Time.now

  ori_dir = Dir.pwd
  Dir.chdir("#{@outdir}/align-genes-pep/")

  is_done = 1
  if Dir["*.pep"].length == Dir["*.aln"].length
    Dir["*.aln"].each do |a|
      if File.size(a) == 0
        is_done = 0
      end
    end
  else
    is_done = 0
  end

  if is_done==0
    Parallel.map(Dir["*.pep"], in_processes: @proc) { |f|
      mafft_align f
    }
  end

  end_time = Time.now
  c_time = Helper.sec2str(end_time-start_time)
  print "done (#{c_time})\n"

  # FIXME ugly hack to find out the reference genome
  ref_id = Dir["#{ori_dir}/#{@genomes_list[0]}/*.pep"][0].split('/')[-1].gsub(".pep","")

  concat_alignments "align-genes-pep.all.fasta", ref_id

  Dir.chdir(ori_dir)

end

#mafft_aln(aln_opt) ⇒ Object 



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# File 'lib/bacterial-comparator.rb', line 376

def mafft_aln aln_opt

  if aln_opt == "both"
    mafft_align_all_pep
    mafft_align_all_dna
  elsif aln_opt == "prot"
    mafft_align_all_pep
  elsif aln_opt == "dna"
    mafft_align_all_dna
  end

end

#raxml_tree(aln_opt, bt) ⇒ Object 



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# File 'lib/bacterial-comparator.rb', line 427

def raxml_tree aln_opt, bt

  if aln_opt == "both"
    raxml_tree_dna bt
    raxml_tree_pep bt
  elsif aln_opt == "prot"
    raxml_tree_pep bt
  elsif aln_opt == "dna"
    raxml_tree_dna bt
  end

end

#raxml_tree_dna(bt) ⇒ Object 



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# File 'lib/bacterial-comparator.rb', line 390

def raxml_tree_dna bt
  print "# Genes DNA tree creation (RAXML).."
  start_time = Time.now
  ori_dir = Dir.pwd
  Dir.chdir(@outdir)
  Dir.mkdir("tree-genes-dna") if ! Dir.exists?("tree-genes-dna")
  current_dir = Dir.pwd
  tree_dir = "#{current_dir}/tree-genes-dna"
  cmd = system("#{@root}/raxml.linux -T #{@proc} -f d -N #{bt} -s align-genes-dna.all.fasta  -m GTRGAMMA -p 123454321 -n DnaTree -w #{tree_dir} > /dev/null 2>&1")
  cmd = system("cat #{tree_dir}/RAxML_result.DnaTree.RUN.* >> #{tree_dir}/RAxML_result.BS")
  cmd = system("#{@root}/raxml.linux -T #{@proc} -f b -z #{tree_dir}/RAxML_result.BS -t #{tree_dir}/RAxML_bestTree.DnaTree -m GTRGAMMA -n DNA_BS_TREE -w #{tree_dir} > /dev/null 2>&1")
  cmd = system("ln -s #{tree_dir}/RAxML_bipartitionsBranchLabels.DNA_BS_TREE #{tree_dir}/../tree-genes-dna.nwk")
  Dir.chdir(ori_dir)
  end_time = Time.now
  c_time = Helper.sec2str(end_time-start_time)
  print "done (#{c_time})\n"
end

#raxml_tree_pep(bt) ⇒ Object 



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# File 'lib/bacterial-comparator.rb', line 408

def raxml_tree_pep bt
  print "# Proteins AA tree creation (RAXML).."
  start_time = Time.now
  ori_dir = Dir.pwd
  Dir.chdir(@outdir)
  Dir.mkdir("tree-genes-pep") if ! Dir.exists?("tree-genes-pep")
  current_dir = Dir.pwd
  tree_dir = "#{current_dir}/tree-genes-pep"
  cmd = system("#{@root}/raxml.linux -T #{@proc} -f d -N #{bt} -s align-genes-pep.all.fasta  -m PROTGAMMAAUTO -p 123454321 -n PepTree -w #{tree_dir} > /dev/null 2>&1")
  cmd = system("cat #{tree_dir}/RAxML_result.PepTree.RUN.* >> #{tree_dir}/RAxML_result.BS")
  cmd = system("#{@root}/raxml.linux -T #{@proc} -f b -z #{tree_dir}/RAxML_result.BS -t #{tree_dir}/RAxML_bestTree.PepTree -m PROTGAMMAAUTO -n PEP_BS_TREE -w #{tree_dir} > /dev/null 2>&1")
  cmd = system("ln -s #{tree_dir}/RAxML_bipartitionsBranchLabels.PEP_BS_TREE #{tree_dir}/../tree-proteins-aa.nwk")
  Dir.chdir(ori_dir)
  end_time = Time.now
  c_time = Helper.sec2str(end_time-start_time)
  print "done (#{c_time})\n"
end

#read_prot_synteny ⇒ Object 



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# File 'lib/bacterial-comparator.rb', line 43

def read_prot_synteny

  print "# Reading genome synteny files - from genome annotations.."
  start_time = Time.now
  synteny = {}
  @genomes_list.each do |g|
    genome_synteny = []
    file = File.open("#{g}/Prot-Synteny.tsv", "r")
    l = file.gets             # skip header
    while l = file.gets
      # AAK98805.1      spr0001 453     1.0     100.0   ABAC01000005_14 453     1.0 1|0
      lA = l.chomp.split("\t")
      synteny[lA[0]] =  [] if ! synteny.has_key? lA[0]
      synteny[lA[0]] << {ref_cov: lA[3].to_f, pId: lA[4].to_f, query_prot: lA[5], query_cov: lA[7].to_f}
      genome_synteny << lA[0]
    end
    @ref_prot.each do |ref_prot|
      if ! genome_synteny.include? ref_prot
        synteny[lA[0]] << {ref_cov: "-", pId: "-", query_prot: "-", query_cov: "-"}
      end
    end
    file.close
  end
  end_time = Time.now
  c_time = Helper.sec2str(end_time-start_time)
  print "done (#{c_time})\n"

  synteny

end