Difference between revisions of "Transgenes"

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[[All_OA_tables#trp_tables_Transgene | trp tables]]
 
[[All_OA_tables#trp_tables_Transgene | trp tables]]
 
*Community submission form
 
*Community submission form
===Form mappings===
+
===Submission Form mappings===
  
 
{| border="1" cellpadding="2" cellspacing="2" {{table}}
 
{| border="1" cellpadding="2" cellspacing="2" {{table}}
| align="center" style="background:#f0f0f0;"|'''uPtag'''
+
| align="center" style="background:#f0f0f0;"|'''Form field label'''
 
| align="center" style="background:#f0f0f0;"|'''OA label'''
 
| align="center" style="background:#f0f0f0;"|'''OA label'''
 
| align="center" style="background:#f0f0f0;"|'''OA pgtable'''
 
| align="center" style="background:#f0f0f0;"|'''OA pgtable'''
 
| align="center" style="background:#f0f0f0;"|'''ontology_type'''
 
| align="center" style="background:#f0f0f0;"|'''ontology_type'''
| align="center" style="background:#f0f0f0;"|'''ontology_table'''
+
| align="center" style="background:#f0f0f0;"|'''ontology_table (source)'''
| align="center" style="background:#f0f0f0;"|'''Term Info'''
+
| align="center" style="background:#f0f0f0;"|'''Term Info to show in OA'''
| align="center" style="background:#f0f0f0;"|'''Gray out for new picture'''
+
| align="center" style="background:#f0f0f0;"|'''Ex values to show on form'''
 
| align="center" style="background:#f0f0f0;"|'''Required field'''
 
| align="center" style="background:#f0f0f0;"|'''Required field'''
 +
| align="center" style="background:#f0f0f0;"|'''notes for Juancarlos'''
 
|-
 
|-
| |Your name||Contact(multiontology on WBPersons)||trp_person||single ontology||WBPerson ontology||e.g.:id : WBPerson1752 name : Hope Dang email : hoped@u.arizona.edu||yes||yes
+
| |Your name||Person)||trp_person||multi-ontology||WBPerson ontology||e.g.:id : WBPerson1752 name : Hope Dang email : hoped@u.arizona.edu|| WBPerson9382 or Monica Duthie||yes||the value entered in this field should show term info on the submission page with the Name, e-mail, affiliation and address. There should also be a link to the person submission form for the person to edit their details. If the person doesn't exist in the database yet, they should be sent to the person submission form as well. That is this form should be restricted to people who have a true WBPersonID
|-
 
| |Your E-mail address||can this be pulled from person tables? ||trp_email needs to be created?||smalltext||-||-||yes||yes
 
 
|-
 
|-
 
| |Gene (e.g. daf-16, ZC132.7)||PGene||exp_gene||ontology on WBGene||gin_tables||as in Expression OA Gene term info e.g.:id :WBGene00007099 locus:trx-2 synonym:B0024.9 synonym:CELE_B0024.9||yes||yes
 
| |Gene (e.g. daf-16, ZC132.7)||PGene||exp_gene||ontology on WBGene||gin_tables||as in Expression OA Gene term info e.g.:id :WBGene00007099 locus:trx-2 synonym:B0024.9 synonym:CELE_B0024.9||yes||yes

Revision as of 23:25, 19 February 2015

back to Caltech documentation

Transgene objects

We curate all integrated (Is, In, Si), and select extrachromosomal (Ex) transgenes.

For each transgene, we extract the following information:

  • name of the transgene*
  • genomic expression summary
  • construct <as of WS245>
  • co-injection markers (as noted as part of the genomic expression summary)
  • method of integration, and to what LG it was mapped, if known
  • which papers use the transgene as part of an experiment
  • other names for the transgene reported by different authors

{*In cases where a transgene has not been assigned a name, or one that does not adhere to standard nomenclature, we assign a name based on the WBPaperID.}


The construct contains the specific information for the array elements

  • plasmid name
  • promoter(s)
  • reporters (GFP, YFP, LacZ, etc.),
  • gene product(s)
  • etc.

Transgene model (WS245)

WormBase_Model:Transgene

?Transgene      Evidence #Evidence
		Public_name UNIQUE ?Text
		Summary UNIQUE ?Text
		Synonym ?Text
		Corresponding_variation UNIQUE ?Variation XREF Corresponding_transgene //put in to unambiguously associate the allele/transgene
		Construction Construct   ?Construct XREF Transgene_construct
     			     Coinjection ?Construct XREF Transgene_coinjection
     			     Coinjection_other ?Text //for coinjection markers that are not specified as a construct
     			     Integration_method UNIQUE ?Text
     			     Integrated_from ?Transgene XREF Transgene_derivation
     			     Laboratory ?Laboratory #Lab_Location
     			     Author ?Author
		Construction_summary ?Text
		Genetic_information Extrachromosomal
				    Integrated
				    Map ?Map  #Map_position  //needed for transgenes with no granular mapping, e.g., just mapped to a LG
                                              Mapping_data 2_point ?2_point_data      //deleted for WS245, rolled back for WS246
				    Map_evidence #Evidence
				    Phenotype ?Phenotype XREF Transgene #Phenotype_info
				    Phenotype_not_observed ?Phenotype XREF Not_in_Transgene #Phenotype_info
		Used_for Transgene_derivation ?Transgene XREF Integrated_from
			 Expr_pattern ?Expr_pattern XREF Transgene
			 Marker_for   ?Text #Evidence
			 Interactor ?Interaction
		Associated_with Marked_rearrangement ?Rearrangement XREF By_transgene
				Strain ?Strain XREF Transgene
		Reference ?Paper XREF Transgene
		Species UNIQUE ?Species
		Remark ?Text #Evidence
                Historical_gene   ?Gene #Evidence

--Kyook (talk) 22:25, 29 December 2014 (UTC)

Transgene first pass:

In/Is/Si transgenes

Arun and Wen have automated the identification of papers that contain transgenes by using Textpresso to scan the C. elegans corpus of papers for the regular expressions (1-3 capital letter)Is, In, or Si (1-4 digits). This script will miss any transgenes that do not have a standard name using fitting this regex.

Transgene names are extracted during a cron job and stored in

http://textpresso-dev.caltech.edu/transgene/transgenes_in_regular_papers.out

"...paper.sup.1" means the transgene name was mentioned in the supplementary file.

Transgene-paper information is picked up by a cron job every Monday at 4am and entered into the trp tables with curator "Arun". Script is found here:

/home/postgres/work/pgpopulation/textpresso/transgene/update_textpresso_transgene.pl

Only the 'Is' and 'Si' lines are entered into postgres. Only paper connections are attached to pre-existing 'In' lines. 'In' is not an accepted nomenclature for transgenes; however, this nomenclature had been used in the past, so there are a handful of transgenes that are valid and should be consistently curated for papers that use report them.

Ex transgenes

Ex transgenes are curated in association with expression patterns, overexpression gene function phenotype, genetic interaction and gene regulation experiments. Although a script was employed to find all Ex transgenes along with genomic summaries in the C. elegans corpus; these have not been consistently curated as a rule. When an extrachromosomal array with a standardized name has been used, it should be curated like all integrated arrays. This output file can be viewed here:

http://textpresso-dev.caltech.edu/transgene/transgenes_summary.out

Dealing with false positive transgenes

False positive transgenes include

  • "In" lines that are chromosomal inversions rather than transgenes.
  • typos in transgene names
  • formatting-induced textpresso extraction errors of transgene names, such as when transgenes are referred to as syIs13-19 in the paper, in which case Textpresso reports it as syIs1319 (i.e., the hyphen disappears sometimes during pdf2Text conversion).

Initially, these transgenes were added to:

/home/acedb/wen/phenote_transgene/ObsoleteTg.txt

which was called on by update_textpreso_transgene.pl to identify objects that shouldn't be entered into the trp tables

All bad transgene objects are now entered into the trp tables and appended with "FAIL". Transgenes that match a pre-existing Transgene public_name-paper association marked as FAIL are ignored during trp table population.

Curation

  • The Textpresso transgene search deposits the transgene name and all new paper instances of the transgene directly into the transgene table.

trp tables

  • Community submission form

Submission Form mappings

Form field label OA label OA pgtable ontology_type ontology_table (source) Term Info to show in OA Ex values to show on form Required field notes for Juancarlos
Your name Person) trp_person multi-ontology WBPerson ontology e.g.:id : WBPerson1752 name : Hope Dang email : hoped@u.arizona.edu WBPerson9382 or Monica Duthie yes the value entered in this field should show term info on the submission page with the Name, e-mail, affiliation and address. There should also be a link to the person submission form for the person to edit their details. If the person doesn't exist in the database yet, they should be sent to the person submission form as well. That is this form should be restricted to people who have a true WBPersonID
Gene (e.g. daf-16, ZC132.7) PGene exp_gene ontology on WBGene gin_tables as in Expression OA Gene term info e.g.:id :WBGene00007099 locus:trx-2 synonym:B0024.9 synonym:CELE_B0024.9 yes yes
Used for Expression Pattern Analysis? Expression-Remark; Picture-Remark exp_remark;pic_remark bigtext - no no
Transgene name Public nameTransgene trp_publicname multiontology trp_tables as in expression OA GO Transgene info: name : WBTransgene00000659 trp_publicname : Is34(eEx85) Paper : WBPaper00003958 trp_summary : [(mab-9::GFP + pCes1943)]. yes transgene OR construct section should have data. try to see if you can grey out construct stuff if transgene is entered
Genotype Summary (e.g.: daf-16::GFP) Construct-Summary cns_summary smalltext when transfer please put it in square brackets - yes yes if no transgene is selected
Construction Details Construct-Construction Details cns_constructionsummary bigtext we should put a question mark next to the name to give more info when clicked - yes yes if no transgene is selected
DNA sequence Construct-DNA cns_dna multi-bigtext joined with pipes - if the DNA sequence, Reporter and Vector fields are filled in, then the submission should generate a construct object and append the cnstID to the expression pattern object yes yes if no transgene is selected
3'UTR Construct-3UTR cns_threeutr multiontology on WBGene gin_tables as in Expression OA Gene term info e.g.:id :WBGene00007099 locus:trx-2 synonym:B0024.9 synonym:CELE_B0024.9 yes no
Reporter Construct-Reporter cns_reporter multiontology values in cns_reporter-Karen is OK in turning the list into an ontology - yes yes if no transgene is selected
Backbone Vector Construct-Clone cns_clone multiontology clone tables as in construct OA clone field: id: D1001 name: "D1001"remark: "C. elegans genomic clone not used in the final reference genome, submitted by The C. elegans Genome Sequencing Consortium."type: "Cosmid" yes yes if no transgene is selected
Fusion Type Construct-Construct Type cns_constructtype ontology right now is not an ontology, Karen said we can turn this in an ontology, too - yes yes if no transgene is selected
Laboratory Construct-Laboratory cns_laboratory multiontology WB laboratory as in Construct OA -> laboratory field: id: LK name: "LK" Mail: "Harvey Mudd College, Claremont CA" Representatives: "Jim Manser"

Allele_designation: "gs"||yes||yes if no transgene is selected

Public Name (e.g.: PJKL491, pCK6) Construct-Public Name cns_publicname smalltext - yes yes
Construct Comments Construct-Remark cns_remark bigtext - yes no
Submit button
Save Button Save button saves to a file loadable by URL, filename WBPerson+Date, sends email to user with Link. Once link is loaded and either Submitted or Saved again, delete the previous save file. If a deleted file tries to get loaded by URL, say it's been submitted or saved over.
Help Button will link to the help page http://www.wormbase.org/tools/support
the fields below are about functionality, not fields that should be on the form
No Dump Transgene-No Dump; Construct-No Dump cns_nodump (needs to be created) all submissions will for now be set as no_dump
Reporter Gene Expression-Type exp_exprtype set to Reporter_gene every submission will have the type set to Reporter_gene
If user fills out the 'new construct' section populate in Exp_OA the exp_construct field exp_construct multiontology
Picture OA -> Expression pattern field populate in Pic_OA the pic_exprpattern field pic_exprpattern multiontology


Curation tools

Transgene .ace dumper

The transgene.ace dumper was written by Juancarlos and Wen to translate the transgene postgres data into .ace format for uploading into AcEDB. The script is located on tazendra. The output file is dumped into the same directory.

Test the .ace in CitaceMinus

  • make sure the file reads in fine.
  • look at all the transgene objects to make sure there are no strange looking ones.
  • do a count of objects before and after the read-in to make sure the number of new objects is reasonable.

A cron job set up by Juancarlos and Wen runs the Transgene .ace dumper script on Thursday mornings at 6am and deposits it on citace at 8am. If there has been any new data or changes in data between testing the file and the Thursday morning dump, make sure to rerum the script and transfer the new data dump to citace for upload.

Requested changes

Changes recorded on https://bitbucket.org/kyook/ky_wbprojects/wiki/transgene_dump_ace.pl

--kjy 23:26, 20 June 2012 (UTC)

  • change dump cron job to Wed morning, spica still calls it at 8am on Thursday
  • fix source of allele codes for obo_laboratory, URL for the lab-allele designations

http://www.cbs.umn.edu/cgc/lab-allele Juancarlos will parse out the page and create a local copy. A cron job will compare the page to the local copy, if the page is altered in format the table will revert to the local copy.


Handling Dead Genes During Dump Process

The dumper script will now (as of May, 2013) run an automatic check for dead genes in any gene field (for transgenes this applies to "Driven_by_gene", "Gene" and "3_UTR" fields/tags). Any genes that are considered dead that are referenced in an Transgene object in the OA will be handled in the following manner:

1) If there is a replacement for the gene (i.e. the gene has merged into another gene), the dead gene will be dumped into a "Historical_gene" field in the .ACE file, the replacement gene will fill the original gene field. A comment will be added to the Historical_gene field via the #Evidence hash. The original gene field (now with the updated gene reference) will be printed with an "Inferred_automatically" tag after the gene. So, for example, if WBGene00001234 is now a dead gene that has been merged into WBGene00002345:

Gene  "WBGene00001234"

becomes

Gene  "WBGene00002345"  Inferred_automatically
Historical_gene  "WBGene00001234"  Remark  "Note: This object originally referred to WBGene00001234.
WBGene00001234 is now considered dead and has been merged into WBGene00002345. WBGene00002345 has 
replaced WBGene00001234 accordingly."

2) If there is no replacement for the gene (Dead or Suppressed), we would dump the following:

Historical_gene  "WBGene00001234"  Remark  "Note: This object originally referred to a gene
 (WBGene00001234) that is now considered dead. Please interpret with discretion."

OR

Historical_gene  "WBGene00001234"  Remark  "Note: This object originally referred to a gene
 (WBGene00001234) that has been suppressed. Please interpret with discretion."

and lastly,

3) If the gene has undergone a split, the interaction will still get dumped, but such genes will be printed out in the error output file of the dumping script for a curator to go back and manually change according to best judgement.


Gene Examples:
A split gene: WBGene00012507
A merged gene: WBGene00007524
A dead gene: WBGene00007814
A suppressed gene: WBGene00015490

Cross curation with other curators

Transgenes are used by other datatypes; Expr_pattern, Phenotype, Gene_regulation, Interaction. All curators that use transgenes will need to run a script before each upload to make sure all their transgenes are valid objects.

Gene_regulation

The gene regulation curator can create new transgenes using the Transgene OA or request them from the transgene curator.

Expr_pattern

The expression pattern curator requires many transgene objects to be created on the fly. Rather than impeding the curation flow, when the expression pattern curator needs a transgene that has not been created already, they enter the relavant information for the transgene in their Reporter gene text box. A script that is manually run will check for lines that have text in the reporter gene field and missing a value in the transgene field. This script will create a new object in the transgene OA containing with the corresponding paper id, expression curator as curator, and the remark field populated with the reporter gene text. A synonym will be created based on the expression pattern value with an appended _Ex. This temporary name will be deposited in the synonym field of the transgene OA for that newly created object. See this wiki page for more information: http://wiki.wormbase.org/index.php/Expression_Pattern#Exporting_Reporter_Gene_description_from_Expr_pattern_OA_to_Transgene_OA

The transgene curator needs to

  • verify that the object created by the expression pattern curator is not a duplicate transgene, if it is a duplicate, the transgene curator will merge the transgene into the preexisting one, this will make the new transgene invalid its information will not be dumped. The new transgene ID and other synonyms will be pipe added to the synonym list of the pre-existing object
  • assign a public name if it exists or if needed
  • fill in all other relevant information

Interaction

The interaction curator can create new transgenes using the Transgene OA or request them from the transgene curator.

Strain

Strain data from the CGC has relevant remarks about the transgene in the strain. Often, there are discrepancies between how an author describes a transgene's genotype and how a CGC strain submitter enters the transgene details. Near the end of every data upload, Mary Ann processes a strain report from the CGC and creates a file transgene_report.<date>.txt that maps transgenes mentioned in the CGC file to WBTransgene IDs, then sends the file to the WB transgene curator.
In the transgene_report, the data shows like:

Transgene : staIs1
Strain : ABR5
Description : staIs1 [pie-1p::GFP + unc-119(+)]. Superficially wild-type. Maintain 
under normal conditions. Reference: This strain is used as the empty vector control 
in Greer EL et al Nature 2010 doi: 10.1038/nature09195.
WBID : WBTransgene00017210

The file needs to be placed on tazendara (note name change) postgres/work/pgpopulation/transgene/transgene_report.txt
Run script populate_maryann_transgene.pl

The populate_maryann_transgene.pl script
1. Enter transgenes from CGC file that are missing from WB.
Start with transgene name, map to WBTransgene ID from trp_name/trp_publicname.

  • If no transgene match, add trp_name (generate trp_name (WBTransgeneID)if it didn't exist already) trp_publicname, trp_strain, trp_cgcremark, and trp_summary.
  • If there is a transgene match only add/change whatever is new/different
    • compare and add strains to trp_strain, pipe separate, ' | ', multiple values, output in out.strains.
nIs407 10031	PG 	CGC MT19635	trp_strain NOW MT19635
nIs408 8234	PG MT20298	CGC MT19756 | MT20298	trp_strain NOW MT19756 | MT20298
nIs425 NEW	PG 	CGC MT19851	trp_strain NOW MT19851

Line 1 no strain existed for pgid 10031 nIs407, the CGC strain was added to trp_strain.
Line 2 different strain existed for pgid 8234 nIs408, CGC strain was merged and pipe separated.
Line 3 nIs425 did not exist in pg, a line was created, and the CGC strain was added to trp_strain.

2. Extract CGC transgene genotype(s), compare it to WB transgene trp_summary. trp_table changes are recorded in out.summary. The transgene should be in square brackets after the transgene name (optional space or words 'contains' or 'is' may be between the transgene name and genotype) in the Description field. There are many cases where the same transgene is described differently in the CGC, the script designates the longest CGC description as the canonical one and pipe separates the remaining genotypes in trp_synonym, with the original trp_summary genotype if it doesn't match the chosen CGC description.

If there is no transgene name mentioned in the Description that matches the Transgene value, pretend there is no genotype, but there is a strain and transgene. Put all unmatched and transgenes with no recognizable genotypes in file cgc_transgene_errors. If the transgene does not exist in pg, a new line, with pgid, trp_publicname, trp_strain, trp_laboratory 'CGC' and trp_curator 'Mary Ann' will be created, but no genotype will be assigned.

Genotype goes to trp_summary, anything that was in summary goes to trp_synonym aggregating with what was there before and separating with ' | '. There can only be one genotype in trp_summary, and it cannot match anything in trp_synonym.

Get the cgcgenotypes
sort them
get the longest one to be the genotype
get the trp_summary values into an aggregate list
get the trp_synonyms values into the aggregate list
get the remaining cgcgenotypes that are not the longest one into the
  aggregate list
if cgcgenotype different from pgsummary, update, and add pgsummary to
  aggregate list
if aggregate list different from pg synonyms, replace trp_summary with
  aggregate list
asIs2 72  	PG [pie-1::mcherry-egg-3]	PGSYN 
		CGC [pie-1::mcherry::egg-3] | [unc-119(+) + ppie-1::GFP::egg-1]
		72	trp_summary NOW [unc-119(+) + ppie-1::GFP::egg-1]
		72	trp_synonym NOW [pie-1::mcherry-egg-3] | [pie-1::mcherry::egg-3]

Line 1 transgene pgid trp_summary trp_synonym.
Line 2 CGC genotypes, aggregated and pipe separated
Line 3 pgid trp_summary replaced by longest CGC genotype
Line 4 pgid trp_synonym aggregate of original trp_summary(PG), original trp_synonyms(PGSYN), and remaining CGC genotypes.

First run was done on all genotypes. In future runs only do it if the genotype is different from the trp_summary.

3. Map WB annotated genes with CGC genotypes, output in out.summary. In many cases, CGC genotypes use a different form of the gene name than what was published. To make it easier to compare the CGC genotypes with WB transgene genotypes, it is helpful to have a gene name mapping output for these elements.
Look at existing trp_gene and trp_drivenbygene, and compare with genes in all gin_locus, gin_synonyms, gin_seqname.
Output on 3 columns which ones matched somewhere in the genotype, which had no match, and whether there was no match at all.

arIs36 45  	PG [phsp::ssGFP]	PGSYN 
				CGC [phsp::ssGFP]
		45	trp_summary NOW [phsp::ssGFP]
		45	trp_synonym NOW 
		45	Matched Gene: None 
		45	Gene Synonyms: None 

asIs2 72  	PG [pie-1::mcherry-egg-3]	PGSYN 
		CGC [pie-1::mcherry::egg-3] | [unc-119(+) + ppie-1::GFP::egg-1]
		72	trp_summary NOW [unc-119(+) + ppie-1::GFP::egg-1]
		72	trp_synonym NOW [pie-1::mcherry-egg-3] | [pie-1::mcherry::egg-3]
		72	Matched Gene: WBGene00004027 pie-1
		72	Matched Gene: WBGene00009701 egg-3
		72	Gene Synonyms: WBGene00004027 CELE_Y49E10.14, Y49E10.14, pic-1
		72	Gene Synonyms: WBGene00009701 CELE_F44F4.2, F44F4.2

axEx1125 78  	PG [pie-1::gfp-mex-5]	PGSYN 
		CGC [pKR2.04 + pRF4 + N2 genomic DNA]
		78	trp_summary NOW [pKR2.04 + pRF4 + N2 genomic DNA]
		78	trp_synonym NOW [pie-1::gfp-mex-5]
		78	Matched Gene: None 
		78	Gene Synonyms: WBGene00003230 CELE_W02A2.7, W02A2.7, mex-5
		78	Gene Synonyms: WBGene00004027 CELE_Y49E10.14, Y49E10.14, pic-1, pie-1

axIs1427 5676  	PG [pie-1prom::LAP::DCAP-1]	PGSYN 
		CGC [pCG26/ LAP-tag DCP-1]
		5676	trp_summary NOW [pCG26/ LAP-tag DCP-1]
		5676	trp_synonym NOW [pie-1prom::LAP::DCAP-1]
		5676	Matched Gene: None 
		5676	Gene Synonyms: WBGene00004027 CELE_Y49E10.14, Y49E10.14, pic-1, pie-1
		5676	Gene Synonyms: WBGene00021929 CELE_Y55F3AM.12, Y55F3AM.12, dcap-1, dcp1


Full entries in out.summary.
For arIs36, the CGC genotype matched trp_summary value, not need to compare genes.
For asIs2, the CGC genotype did not match trp_summary value, genes within old and new trp_summary values need comparing. Matched genes are noted. Gene synonyms are supplied.
For axEx1125, trp_summary replaced, but no genes match, no gene or gene synonym recognized in CGC genotype, this transgene needs resolving.
For axIs1427, trp_summary replaced, gene synonyms match for DCAP-1 and DCP-1.

4. errors file in output are for pg entries that can't be compared, for example if there are duplicate transgene public_names, and neither of them have been labeled as 'FAIL'. These objects will need to be resolved or the script will break.

5. All changes to the postgres tables are listed in out.pg. Use these grep searches to display the number of changes to the different tables:

>grep " trp_strain VALUES " out.pg | wc -l
902
>grep " trp_summary VALUES " out.pg | wc -l
4450
>grep " trp_synonym VALUES " out.pg | wc -l
4396
>grep " trp_publicname VALUES " out.pg | wc -l
55

Other tables that can be queried from the out.pg are:

  • trp_summary_hst
  • trp_strain_hst
  • trp_synonym_hst
  • trp_publicname_hst NOTE: trp_publicname and trp_publicname_hst table change when there is a new entry
  • trp_laboratory NOTE: trp_laboratory and trp_laboratory_hst table change when there is a new entry
  • trp_laboratory_hst

NOTE: No diff file necessary between transgene_reports, always run on everything, only do stuff when it's new.

Web Displays

Page for mapped transgenes http://wiki.wormbase.org/index.php/Mapped_Transgenes