Open ramadatta opened 7 years ago
Hi @ramadatta, Can I have first few lines from your GFF3 input file?
Hi Vipin,
Please find it here:
##gff-version 3
#!gff-spec-version 1.21
#!processor NCBI annotwriter
#!genome-build ASM185804v2
#!genome-build-accession NCBI_Assembly:GCF_001858045.1
#!annotation-date 5 December 2016
#!annotation-source NCBI Oreochromis niloticus Annotation Release 103
##sequence-region NC_031965.1 1 38372991
##species https://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?id=8128
NC_031965.1 RefSeq region 1 38372991 . + . ID=id0;Dbxref=taxon:8128;Name=LG1;dev-stage=adult;gbkey=Src;genome=chromosome;isolate=F11D_XX;linkage-group=LG1;mol_type=genomic DNA;sex=female;tissue-type=blood
NC_031965.1 Gnomon gene 11049 13118 . - . ID=gene0;Dbxref=GeneID:102075536;Name=LOC102075536;gbkey=Gene;gene=LOC102075536;gene_biotype=lncRNA
NC_031965.1 Gnomon lnc_RNA 11049 13118 . - . ID=rna0;Parent=gene0;Dbxref=GeneID:102075536,Genbank:XR_266817.3;Name=XR_266817.3;gbkey=ncRNA;gene=LOC102075536;model_evidence=Supporting evidence includes similarity to: 100%25 coverage of the annotated genomic feature by RNAseq alignments%2C including 13 samples with support for all annotated introns;product=uncharacterized LOC102075536;transcript_id=XR_266817.3
NC_031965.1 Gnomon exon 13071 13118 . - . ID=id1;Parent=rna0;Dbxref=GeneID:102075536,Genbank:XR_266817.3;gbkey=ncRNA;gene=LOC102075536;product=uncharacterized LOC102075536;transcript_id=XR_266817.3
NC_031965.1 Gnomon exon 12125 12153 . - . ID=id2;Parent=rna0;Dbxref=GeneID:102075536,Genbank:XR_266817.3;gbkey=ncRNA;gene=LOC102075536;product=uncharacterized LOC102075536;transcript_id=XR_266817.3
NC_031965.1 Gnomon exon 11049 11689 . - . ID=id3;Parent=rna0;Dbxref=GeneID:102075536,Genbank:XR_266817.3;gbkey=ncRNA;gene=LOC102075536;product=uncharacterized LOC102075536;transcript_id=XR_266817.3
NC_031965.1 Gnomon gene 31634 150960 . - . ID=gene1;Dbxref=GeneID:100690862;Name=LOC100690862;gbkey=Gene;gene=LOC100690862;gene_biotype=protein_coding
Prakki Rama
@ramadatta the input gff3 file lines looks good to me. From the above error message, I am seeing little difference the current version of the code:
https://github.com/vipints/GFFtools-GX/blob/master/gff_to_bed.py#L87
. Can you test with this recent version of the script.
Hi Vipin,
Thank you for quick replies. I have got the gff3 file from NCBI. The following is the result when I used the latest version of your script:
$ python gff_to_bed_latest.py ref_ASM185804v2_top_level.gff3 >out3.bed
Traceback (most recent call last):
File "gff_to_bed_latest.py", line 112, in <module>
__main__()
File "gff_to_bed_latest.py", line 109, in __main__
writeBED(Transcriptdb)
File "gff_to_bed_latest.py", line 66, in writeBED
score = ent1['transcript_score'][idx] if ent1['transcript_score'].any() else score ## getting the transcript score
ValueError: no field of name transcript_score
@ramadatta can you provide me the ncbi link where you downloaded the gff3 file.
Hi Vipin,
Here is the annotation file:
ftp://ftp.ncbi.nlm.nih.gov/genomes/Oreochromis_niloticus/GFF/ref_ASM185804v2_top_level.gff3.gz
Hi @ramadatta,
I have downloaded the file from the above link. There are many features available in this GFF file.
cut -f 2,3 ref_ASM185804v2_top_level.gff3 | sort | uniq -c
The file looks good with the features available. Now I found an issue with feature mappings in the file:
NW_017615909.1 Gnomon gene 431704 433832 . + . ID=gene36112;Dbxref=GeneID:100700432;Name=LOC100700432;gbkey=Gene;gene=LOC100700432;gene_biotype=pseudogene;pseudo=true
NW_017615909.1 Gnomon exon 431704 431773 . + . ID=id778571;Parent=gene36112;Dbxref=GeneID:100700432;gbkey=exon;gene=LOC100700432;model_evidence=Supporting evidence includes similarity to: 4 Proteins
NW_017615909.1 Gnomon exon 433129 433832 . + . ID=id778572;Parent=gene36112;Dbxref=GeneID:100700432;gbkey=exon;gene=LOC100700432;model_evidence=Supporting evidence includes similarity to: 4 Proteins
The exon feature is directly mapped to gene, in this mapping there missing a additional layer of mapping exon -> transcript -> gene. I would say as per the GFF specification even if it is a single transcript gene, the mapping description should go as mentioned before. (gene -> transcript -> exon). May be it is good to contact the genome annotation center (ncbi) where you downloaded the file and inform them that the mapping in GFF is not valid.
in BED format we are representing the transcript feature in a single line.
I could do something that I can kick out these non properly matched annotation features. But I m not sure whether this will clean all single exon gene annotation from this file. I think then the resulting bed format will not contain any single exon gene. That will not be a good fix to this problem. Also you cannot define the missing transcript status whether a mRNA, lnRNA, or any other type of RNA feature, this has to be defined by the annotation center.
Hi @vipints
That appears true. Some transcripts are missing the status. Any other possible solution to think. May be I will try to approach the NCBI annotation team about this. Thanks so much for your help!
Hi Vipin,
Thank you very much for the GFFtools scripts. However, when I run the command,
python gff_to_bed.py ref.gff3 > out2.bed
while converting gff3 to bed 12 format, I receive an error below:
Could I please know how to fix this error?