require 'rubygems' require 'bio' require 'snp_db_models' #establish_connection def guess_sequence_format(reference_genome) file_extension = File.extname(reference_genome).downcase file_format = nil case file_extension when ".gbk", ".genbank", ".gb" file_format = :genbank when ".embl", ".emb" file_format = :embl end return file_format end # A method to populate the strain names in the Strain table. strain_names is an array of strain names. def populate_strains(strain_names) strain_names.each do |strain| s = Strain.new s.name = strain s.save end end # A method to populate the database with the features (genes etc) and the annotations from the embl file. # We include all features that are not 'source' or 'gene' as they are repetitive info. 'CDS' is the gene. # The annotation table includes also the start and end coordinates of the CDS. The strand is also included. the 'locations' method is defined in bioruby under genbank. It must be required at the top (bio). # Also, the qualifier and value are extracted from the embl file and added to the database. def populate_features_and_annotations(sequence_file) sequence_file.features.each do |feature| unless feature.feature == "source" || feature.feature == "gene" db_feature = Feature.new db_feature.start = feature.locations.first.from db_feature.end = feature.locations.first.to db_feature.strand = feature.locations.first.strand db_feature.name = feature.feature db_feature.save puts "populated #{db_feature.name}, start: #{db_feature.start}, end: #{db_feature.end}, strand: #{db_feature.strand} for feature: #{db_feature.id}" # Populate the Annotation table with qualifier information from the genbank file feature.qualifiers.each do |qualifier| a = Annotation.new a.qualifier = qualifier.qualifier a.value = qualifier.value a.save db_feature.annotations << a puts "populated #{a.qualifier} for feature: #{db_feature.id}" end end end end #This method populates the rest of the information, i.e. SNP information, Alleles and Genotypes. # It requires the strain_names as array and the output (vcf file) from mpileup-snp identification algorithm. def populate_snps_alleles_genotypes(strain_names, vcf_file, cuttoff_snp, cuttoff_genotype) strains = Array.new strain_names.each do |strain_name| strain = Strain.find_by_name(strain_name) # equivalent to Strain.find.where("strains.name=?", strain_name).first strains << strain end # open vcf file and parse each line File.open(vcf_file) do |f| # header names header = f.gets header2 = f.gets.chomp column_headings = header2.split("\t") sample_names = column_headings[9..-1] good_snps = 0 # start parsing snps while line = f.gets details = line.split("\t") ref = details[0] ref_pos = details[1] ref_base = details[3] snp_base = details[4] snp_qual = details [5] samples = details[9..-1] genotypes = samples.map do |s| pl, gt, gq = s.chomp.split(":") gt end genotypes_qualities = samples.map do |s| pl, gt, gq = s.chomp.split(":") gq end high_quality_variant_genotypes = Array.new # this will be filled with the indicies of genotypes that are "1/1" and have a quality >= 30 variant_genotypes = Array.new genotypes.each_with_index do |gt, index| if gt == "1/1" variant_genotypes << index if genotypes_qualities[index].to_i >= cuttoff_genotype high_quality_variant_genotypes << index end end end if snp_qual.to_i >= cuttoff_snp && genotypes.include?("1/1") && ! high_quality_variant_genotypes.empty? && high_quality_variant_genotypes.size == variant_genotypes.size # first condition checks the overall quality of the SNP is >=90, second checks that at least one genome has the 'homozygous' 1/1 variant type with quality >= 30 and informative SNP if genotypes.include?("0/0") && !genotypes.include?("0/1") # exclude SNPs which are all 1/1 i.e something strange about ref and those which have confusing heterozygote 0/1s good_snps +=1 # puts good_snps #create snp s = Snp.new s.ref_pos = ref_pos s.save puts "Adding Reference SNP position: #{ref_pos}" # create ref allele ref_allele = Allele.new ref_allele.base = ref_base ref_allele.snp = s ref_allele.save puts "Adding Reference SNP base: #{ref_base}" s.reference_allele = ref_allele s.save # create snp allele snp_allele = Allele.new snp_allele.base = snp_base snp_allele.snp = s snp_allele.save puts "Adding SNP base: #{snp_base}" genotypes.each_with_index do |gt, index| genotype = Genotype.new genotype.strain = strains[index] puts index if strains[index].nil? # print "#{gt}(#{genotypes_qualities[index]}) " if gt == "0/0" # wild type genotype.allele = ref_allele elsif gt == "1/1" # snp type genotype.allele = snp_allele else puts "Strange SNP #{gt}" end genotype.save end end end end end #Here we link the features to snps. Snp.all.each do |snp| x = Feature.where("features.start <= ? AND features.end >= ?", snp.ref_pos, snp.ref_pos).first snp.feature = x snp.save end end