内容简介:Blast比对软件大概是是短序列局部比对软件中最常用的一个了,但是其参数众多,一些参数一直没好好仔细研究过,如下:blast的其实我们一般常用的就是
Blast比对软件大概是是短序列局部比对软件中最常用的一个了,但是其参数众多,一些参数一直没好好仔细研究过,如下:
增加blast比对结果信息
blast的 -outfmt
参数,使 blastp -help
即可查看每个输出格式的信息,如下所示:
*** Formatting options -outfmt <String> alignment view options: 0 = Pairwise, 1 = Query-anchored showing identities, 2 = Query-anchored no identities, 3 = Flat query-anchored showing identities, 4 = Flat query-anchored no identities, 5 = BLAST XML, 6 = Tabular, 7 = Tabular with comment lines, 8 = Seqalign (Text ASN.1), 9 = Seqalign (Binary ASN.1), 10 = Comma-separated values, 11 = BLAST archive (ASN.1), 12 = Seqalign (JSON), 13 = Multiple-file BLAST JSON, 14 = Multiple-file BLAST XML2, 15 = Single-file BLAST JSON, 16 = Single-file BLAST XML2, 18 = Organism Report Options 6, 7 and 10 can be additionally configured to produce a custom format specified by space delimited format specifiers. The supported format specifiers are: qseqid means Query Seq-id qgi means Query GI qacc means Query accesion qaccver means Query accesion.version qlen means Query sequence length sseqid means Subject Seq-id sallseqid means All subject Seq-id(s), separated by a ';' sgi means Subject GI sallgi means All subject GIs sacc means Subject accession saccver means Subject accession.version sallacc means All subject accessions slen means Subject sequence length qstart means Start of alignment in query qend means End of alignment in query sstart means Start of alignment in subject send means End of alignment in subject qseq means Aligned part of query sequence sseq means Aligned part of subject sequence evalue means Expect value bitscore means Bit score score means Raw score length means Alignment length pident means Percentage of identical matches nident means Number of identical matches mismatch means Number of mismatches positive means Number of positive-scoring matches gapopen means Number of gap openings gaps means Total number of gaps ppos means Percentage of positive-scoring matches frames means Query and subject frames separated by a '/' qframe means Query frame sframe means Subject frame btop means Blast traceback operations (BTOP) staxid means Subject Taxonomy ID ssciname means Subject Scientific Name scomname means Subject Common Name sblastname means Subject Blast Name sskingdom means Subject Super Kingdom staxids means unique Subject Taxonomy ID(s), separated by a ';' (in numerical order) sscinames means unique Subject Scientific Name(s), separated by a ';' scomnames means unique Subject Common Name(s), separated by a ';' sblastnames means unique Subject Blast Name(s), separated by a ';' (in alphabetical order) sskingdoms means unique Subject Super Kingdom(s), separated by a ';' (in alphabetical order) stitle means Subject Title salltitles means All Subject Title(s), separated by a '<>' sstrand means Subject Strand qcovs means Query Coverage Per Subject qcovhsp means Query Coverage Per HSP qcovus means Query Coverage Per Unique Subject (blastn only)
其实我们一般常用的就是 -outfmt 5
或者 -outfmt 6
,前者输出XML格式,后者输出TAB分割格式;前者在早期一篇博文Blast+ xml格式解读中提起过(信息比较全,用处也相对比较广),而后者则是平时最为常用的格式(也是一些软件喜欢调用的格式)
TAB格式每列信息如下(可以对照上面的说明理解一下):
qseqid sseqid pident length mismatch gapopen qstart qend sstart send evalue bitscore
但我们有时想要的并不止上述12列信息,比如我还想知道比对结果的覆盖度信息(qcovs:Query Coverage Per Subject)
其实只要在blast比对命令中先事先加上需要增加的列ID即可,如在 outfmt 6
基础上加上覆盖度信息:
-outfmt "6 qseqid sseqid pident length mismatch gapopen qstart qend sstart send evalue bitscore qcovs"
注:需要几列就一直往上加即可,空格分割
分割NR子库
之前分割NR子库选用的是早期一篇博文 创建NR子库以及从NR库提取特定物种分类的序列
但是现在NCBI出了blast-2.8版本,其可支持用NCBI自带代码分割的NR子库的索引作为比对的库,使用比较方便
- Support for a new version of the BLAST database that allows you to limit search by taxonomy as well some other improvements.
当然如果用这个版本的话,NR库也要重新下载了 ftp://ftp.ncbi.nlm.nih.gov/blast/db/v5/
使用方式也比较简单(至少比之前的方法方便了),如果只想比对单一物种(如人:9606的话),命令如下:
blastp –db nr –query query.fasta –taxids 9606 –outfmt 6 –out blast.outfm6
如果想比对NR子库哺乳动物的话,需要先建个哺乳动物子库索引
get_species_taxids.sh -t 40674 > 40674.txids
然后再将序列比对至NR哺乳动物子库
blastp –db nr –query query.fasta –taxidlist 40674.txids –outfmt 6 –out blast.outfm6
具体说明可看: https://ftp.ncbi.nlm.nih.gov/blast/db/v5/blastdbv5.pdf
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以上所述就是小编给大家介绍的《Blast+ 使用补充笔记》,希望对大家有所帮助,如果大家有任何疑问请给我留言,小编会及时回复大家的。在此也非常感谢大家对 码农网 的支持!
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