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Heat shock induced excision of selectable marker genes in transgenic banana by the Cre-lox site-specific recombination system

Paper ID Volume ID Publish Year Pages File Format Full-Text
23589 43454 2012 9 PDF Available
Title
Heat shock induced excision of selectable marker genes in transgenic banana by the Cre-lox site-specific recombination system
Abstract

Selectable marker genes are indispensable for efficient production of transgenic events, but are no longer needed after the selection process and may cause public concern and technological problems. Although several gene excision systems exist, few have been optimized for vegetatively propagated crops. Using a Cre-loxP auto-excision strategy, we obtained transgenic banana plants cv. Grande Naine (Musa AAA) devoid of the marker gene used for selection. We used T-DNA vectors with the cre recombinase gene under control of a heat shock promoter and selectable marker gene cassettes placed between two loxP sites in direct orientation, and a gene of interest inserted outside of the loxP sites. Heat shock promoters pGmHSP17.6-L and pHSP18.2, from soybean and Arabidopsis respectively, were tested. A transient heat shock treatment of primary transgenic embryos was sufficient for inducing cre and excising cre and the marker genes. Excision efficiency, as determined by PCR and Southern hybridization was 59.7 and 40.0% for the GmHSP17.6-L and HSP18.2 promoters, respectively. Spontaneous excision was not observed in 50 plants derived from untreated transgenic embryos. To our knowledge this is the first report describing an efficient marker gene removal system for banana. The method described is simple and might be generally applicable for the production of marker-free transgenic plants of many crop species.

► Selectable marker genes are needed for generating transgenic banana, but are ideally absent from the final transgenic plant. ► We developed an efficient method to excise the marker gene from the genome of transgenic embryos after the selection step. ► The marker removal system is based on the site specific recombinase Cre driven by a heat shock promoter. ► Absence of the selectable marker from transgenic banana was shown by PCR, Southern hybridization and phenotypic analyses. ► Marker removal does not reduce the transformation frequency nor does it lengthen the transformation procedure.

Keywords
Agrobacterium; Banana; Cre-lox; Marker gene excision; Musa; Transgenic plant
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Heat shock induced excision of selectable marker genes in transgenic banana by the Cre-lox site-specific recombination system
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 159, Issue 4, 30 June 2012, Pages 265–273
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us