Intron-mediated improvement of a selectable marker gene for plant transformation using Agrobacterium tumefaciens
Wang, Ming-Bo, Upadhyaya, N. M., Brettell, R. I. S., & Waterhouse, Peter M. (1997) Intron-mediated improvement of a selectable marker gene for plant transformation using Agrobacterium tumefaciens. Journal of Genetics and Breeding, 51(4), pp. 325-334.
In binary vectors, the antibiotic resistance gene used for selection of transformed plant cells is also usually expressed in the transforming Agrobacterium cells. This expression gives the bacterium antibiotic resistance, an unnecessary advantage on selective medium containing the antibiotic. Insertion of a castor bean catalase-1 (CAT-1) gene intron or a Parasponia andersonii haemoglobin gene intron into the coding region of the selectable marker gene, hph, completely abolished the expression of the gene in Agrobacterium, rendering it susceptible to hygromycin B. Use of these modified binary vectors minimized the overgrowth of Agrobacterium during plant transformation. Both of the introns were correctly spliced in plant cells and significantly enhanced hph gene expression in transformed rice tissue. The presence of these introns in the hph coding sequence not only maintained the selection efficiency of the hph gene, but with the CAT-1 intron also substantially increased the frequency of rice transformation. Transgenic lines with an intron-hph gene generally contained fewer gene copies and produced substantially more mRNA of the predicted size. Our results also indicate that transgenic plants with many copies of the transgene were more likely to show gene silencing than plants with 1-3 copies.
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|Item Type:||Journal Article|
|Keywords:||Enhancement, Hph, Hygromycin resistance, Intron insertion, Rice, Transgene, antibiotic resistance, catalase 1, gene silencing, gene targeting, gene transfer, genetic improvement, genetic marker, intron, transgenic plant, Agrobacterium tumefaciens, Parasponia andersonii|
|Subjects:||Australian and New Zealand Standard Research Classification > BIOLOGICAL SCIENCES (060000) > GENETICS (060400)
Australian and New Zealand Standard Research Classification > BIOLOGICAL SCIENCES (060000) > PLANT BIOLOGY (060700)
|Divisions:||Current > Schools > School of Earth, Environmental & Biological Sciences
Current > QUT Faculties and Divisions > Science & Engineering Faculty
|Copyright Owner:||Copyright 1997 L' Unita di Ricerca per la Valorizzazione Qualitativa dei Cereali (C R A - Q C E)|
|Deposited On:||13 Jan 2014 01:53|
|Last Modified:||13 Jan 2014 01:53|
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