使用MCScanX和JCVI共线性区块分析

首先从基因组数据库下载目标物种的全基因组文件和gff3文件
对gff3文件预处理得到对应格式的gff和bed文件

MCScanX:perl -e 'while (<>) { if (m/^(\S+)\t.*\tgene\t(\d+)\t(\d+).*ID=(darer\d+)/) { print "$1\t$4\t$2\t$3\n" } }' darer.genome.gff3 > darer.gff
jcvi:perl -e 'while (<>) { if (m/^(\S+)\t.*\tgene\t(\d+)\t(\d+)\t(\S+)\t(\S+).*ID=(darer\d+)/) { print "$1\t$2\t$3\t$6\t$4\t$5\n" } }' darer.genome.gff3 > zebr.bed

对NCBI的GFF3文件再进行提取,若基因含有可变剪接,则仅保留CDS长度最长的转录本并获得蛋白序列,该过程参考网上其他教程

cat darer.gff gc.gff > all.gff
cat darer.pep.fasta gc.pep.fasta > all.pep.fasta
makeblastdb -in all.pep.fasta -dbtype prot -title all -parse_seqids -out all -logfile all.log
blast.pl blastp all all.pep.fasta 1e-10 160 blast 6
mkdir MCScanx
mv ../blast.tab all.blast
mv ../all.gff ./
MCScanX all
cd ..
mkdir jcvi && cd jcvi
mv ../*.bed ./
mv ../darer.pep.fasta darer.pep
mv ../gc.pep.fasta gc.pep
python -m jcvi.compara.catalog ortholog --dbtype prot --no_strip_names darer gc
python -m jcvi.compara.synteny screen --minspan=30 --simple darer.gc.anchors darer.gc.anchors.new

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