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A point mutation in the Medicago sativa GSA gene provides a novel, efficient, selectable marker for plant genetic engineering

Paper ID Volume ID Publish Year Pages File Format Full-Text
23846 43477 2011 6 PDF Available
Title
A point mutation in the Medicago sativa GSA gene provides a novel, efficient, selectable marker for plant genetic engineering
Abstract

Bacterial selectable marker genes (SMG) conferring antibiotic resistance are valuable tools in plant genetic engineering, but public concern and regulatory requirements have stimulated the development of alternative selection systems. We have previously demonstrated that a mutated Synechococcus elongatus HemL gene encoding glutamate 1-semialdehyde aminotransferase (GSA) is an efficient SMG in alfalfa. In fact, GSA is irreversibly inhibited by gabaculine (3-amino-2,3-dihydrobenzoic acid), but the mutated enzyme is gabaculine insensitive. With the aim to develop a plant derived SMG, we cloned and sequenced the Medicago sativa GSA cDNA and reproduced one of the two mutations associated with gabaculine resistance in Synechococcus, a transversion resulting in a methionine to isoleucine (M → I) substitution. This mutated gene was assessed as a SMG in tobacco and alfalfa Agrobacterium transformation, in comparison with the wild type gene. In tobacco, about 43% of the leaf explants produced green shoots, whereas in alfalfa 47% of the explants produced green embryos in the presence of 30 μM gabaculine when the M → I GSA was introduced. Escapes were absent in tobacco and only 6% in alfalfa. No effect on the plant phenotype was noticed. We propose this new SMG as a widely acceptable alternative to those currently used.

► We have previously demonstrated that a mutated Synechococcus elongatus HemL gene encoding glutamate 1-semialdehyde aminotransferase (GSA) is an efficient selectable marker gene (SMG) in alfalfa by conferring resistance to gabaculine. ► With the aim to develop a plant derived SMG, we cloned and sequenced the Medicago sativa GSA cDNA and introduced a point mutation. ► In both alfalfa and tobacco the new SMG was very efficient and no effect on the plant phenotype was noticed. ► We propose this new SMG as a widely acceptable alternative to those currently used.

Keywords
Alfalfa; Gabaculine; GMO; Selectable marker genes; Tobacco
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A point mutation in the Medicago sativa GSA gene provides a novel, efficient, selectable marker for plant genetic engineering
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 156, Issue 2, 10 November 2011, Pages 147–152
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