[Gmod-help] Problems with Bio::DB::SeqFeature::Store and gff3

Smithies, Russell Russell.Smithies at agresearch.co.nz
Thu Sep 11 22:10:30 EDT 2008


I've spent all morning trying to get gff3 data to display after loading into a SeqFeature database and I'm getting nowhere  :-(
We installed the fresh GBrowse 1.69 release yesterday and had to upgrade to bioperl-live as the version of Bioperl we were using (also the latest release) was too low according to the install script.

Our old GBrowse instances using gff2 data and MySQL are still running fine so I assume it's not BioPerl that's broken.
But when I try to search for anything on our new gff3 GBrowse, I get the usual "landmark maned foobar is not recognized" on the webpage.
The error logs are full of this rather cryptic message:
-------------------------------------------------
[Fri Sep 12 13:59:38 2008] [error] [client 147.158.129.160] Bio::Graphics::BrowserConfig=HASH(0x1742be30) at /usr/lib64/perl5/site_perl/5.8.8/x86_64-linux-thread-multi/Bio/Graphics/Browser.pm line 587., referer: http://devgbrowse.agresearch.co.nz/cgi-bin/gbrowse/dev_oar/?reset=1
[Fri Sep 12 13:59:38 2008] [error] [client 147.158.129.160] Bio::Graphics::BrowserConfig=HASH(0x1742be30) at /usr/lib64/perl5/site_perl/5.8.8/x86_64-linux-thread-multi/Bio/Graphics/Browser.pm line 587., referer: http://devgbrowse.agresearch.co.nz/cgi-bin/gbrowse/dev_oar/?reset=1
[Fri Sep 12 13:59:38 2008] [error] [client 147.158.129.160] Bio::Graphics::BrowserConfig=HASH(0x1742be30) at /usr/lib64/perl5/site_perl/5.8.8/x86_64-linux-thread-multi/Bio/Graphics/Browser.pm line 587., referer: http://devgbrowse.agresearch.co.nz/cgi-bin/gbrowse/dev_oar/?reset=1
[Fri Sep 12 13:59:38 2008] [error] [client 147.158.129.160] Bio::Graphics::BrowserConfig=HASH(0x1742be30) at /usr/lib64/perl5/site_perl/5.8.8/x86_64-linux-thread-multi/Bio/Graphics/Browser.pm line 587., referer: http://devgbrowse.agresearch.co.nz/cgi-bin/gbrowse/dev_oar/?reset=1
[Fri Sep 12 13:59:38 2008] [error] [client 147.158.129.160] Use of uninitialized value in split at /var/www/gbrowse/cgi-bin/gbrowse line 1682., referer: http://devgbrowse.agresearch.co.nz/cgi-bin/gbrowse/dev_oar/?reset=1
-------------------------------------------------

I've kept my config to a minimum to try and debug:

-------------------------------------------------
[GENERAL]
description   = DEVELOPMENT OAR true chromosomes

db_adaptor    = Bio::DB::SeqFeature::Store
db_args       = -adaptor DBI::mysql
                -dsn     dbi:mysql:database=dev_oar_chromosomes;host=gbrowsemysql
user          = username
pass         = password

initial landmark = OAR1:76900..77100

# examples to show in the introduction
examples = OAR1:100..500 XM_001256510 XM_001250379

# Web site configuration info
stylesheet  = /gbrowse/gbrowse.css
buttons     = /gbrowse/images/buttons
tmpimages   = /gbrowse/tmp

[TRACK DEFAULTS]
glyph         = generic
height        = 8
fgcolor       = black
font2color    = gray
label density = 800
bump density  = 10000
link_target	  = _blank


[Test]
feature     = mRNA
glyph       = cds
description = 1
key         = Test
citation    = test
------------------------------------------------------------------------------

And the gff3 data looks like this:


------------------------------------------------------------------------------
##gff-version 3
OAR1	.	chromosome	1	80000	.	+	.	Sequence OAR1
OAR1	btref	mRNA	2988	20491	3756	+	.	ID=XM_001256510;Target=XM_001256510+1+2278;Note=PREDICTED:+Bos+taurus+hypothetical+protein+LOC767882+(LOC767882)%2C+partial+mRNA;Entrez_Gene=767882;GO=GO:0003723;GO_Term=RNA+binding
OAR1	btref	exon	2988	3049	3756	+	.	Parent=XM_001256510;Target=XM_001256510+1+62
OAR1	btref	exon	4092	4204	3756	+	.	Parent=XM_001256510;Target=XM_001256510+63+175
OAR1	btref	exon	6060	6219	3756	+	.	Parent=XM_001256510;Target=XM_001256510+176+335
OAR1	btref	exon	7066	7175	3756	+	.	Parent=XM_001256510;Target=XM_001256510+440+549
OAR1	btref	exon	11913	12119	3756	+	.	Parent=XM_001256510;Target=XM_001256510+550+756
OAR1	btref	exon	12367	12582	3756	+	.	Parent=XM_001256510;Target=XM_001256510+757+972
OAR1	btref	exon	13003	13209	3756	+	.	Parent=XM_001256510;Target=XM_001256510+973+1179
OAR1	btref	exon	13333	13440	3756	+	.	Parent=XM_001256510;Target=XM_001256510+1180+1287
OAR1	btref	exon	14246	14350	3756	+	.	Parent=XM_001256510;Target=XM_001256510+1288+1392
OAR1	btref	exon	14626	14704	3756	+	.	Parent=XM_001256510;Target=XM_001256510+1393+1471
OAR1	btref	exon	15944	16084	3756	+	.	Parent=XM_001256510;Target=XM_001256510+1472+1612
OAR1	btref	exon	16735	16748	3756	+	.	Parent=XM_001256510;Target=XM_001256510+1618+1631
OAR1	btref	exon	16751	16834	3756	+	.	Parent=XM_001256510;Target=XM_001256510+1633+1716
OAR1	btref	exon	16956	17069	3756	+	.	Parent=XM_001256510;Target=XM_001256510+1717+1830
OAR1	btref	exon	17844	17930	3756	+	.	Parent=XM_001256510;Target=XM_001256510+1831+1917
OAR1	btref	exon	20262	20491	3756	+	.	Parent=XM_001256510;Target=XM_001256510+2049+2278
OAR1	btref	mRNA	76974	77060	174	-	.	ID=XM_001250379;Target=XM_001250379+10+100;Note=PREDICTED:+Bos+taurus+similar+to+FARP2+protein+(LOC781869)%2C+mRNA;Entrez_Gene=781869
OAR1	btref	exon	76974	76977	174	-	.	Parent=XM_001250379;Target=XM_001250379+97+100
OAR1	btref	exon	76978	77053	174	-	.	Parent=XM_001250379;Target=XM_001250379+20+95
OAR1	btref	exon	77054	77060	174	-	.	Parent=XM_001250379;Target=XM_001250379+10+16
---------------------------------------------------------------------------


As far as I can see, the database is loading correctly:

----------------------------------------------
[smithiesr at impala oar]$ bp_seqfeature_load.pl  --create --verbose --fast --user username --password password  -dsn 'dbi:mysql:host=gbrowsemysql;database=dev_oar_chromosomes' dev_gff/OAR1.test.gff3
loading dev_gff/OAR1.test.gff3...

Building object tree... 0.00s
Loading bulk data into database... 0.01s
load time:  0.07s
------------------------------------------------

Any ideas what to try next??
Have I overlooked something obvious?


Thanx,


Russell Smithies 

Bioinformatics Applications Developer 
Invermay  Research Centre 
Puddle Alley, 
Mosgiel, 
New Zealand 
www.agresearch.co.nz 





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