Construction of Plant Expression Constructs Harboring Full-length Bt Cry1Ac22 Toxin Gene and Truncated Functional Domains of Bt Cry1Ac22 Toxin and Arabidopsis Transformation
1. Hainan Institute of Tropical Agricultural Resources, Sanya, 572025, P.R. China
2. Alkali Soil Natural Environmental Science Center (ASNESC), Northeast Forestry Uinversity, Harbin, 150040, P.R. China
3. College of Life and Technology Science, Guangxi University, Nanning, 530004, P.R. China
Bioscience Methods, 2011, Vol. 2, No. 3 doi: 10.5376/bm.2011.02.0003
Received: 27 Dec., 2010 Accepted: 19 May, 2011 Published: 30 May, 2011
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Preferred citation for this article:
Liu et al., 2011, Construction of Plant expression Constructs Harboring Full-length Bt Cry1Ac22 Toxin Gene and Truncated Functional Domains of Bt Cry1Ac22 Toxin and Arabidopsis Transformation, Bioscience Methods, Vol.2 No.2 (doi:10.5376/bm.2011.02.0003)
The full-length Bt toxin gene (3 534 bp) and truncated functional domains of Bt toxin gene (1 959 bp) of cry1Ac22 were amplified by PCR from Bt strain W015-1. The PCR products were ligated into plant expression vector pBI121 cutting off the ß-glucuronidase gene to make the constructs of pBI121-Cry1Ac22F and pBI121-Cry1Ac22T. The constructs carrying with Kanamycin resistant marker were transferred into T-DNA vector and then validated by restricted enzyme digestion and PCR identification. Arabidopsis transformation with the transfered T-DNA vector were performed during the flowering stage mediated by Agrobacterium tumufaciens. Transgenic Arabidopsis seeds with positive Kanamycin resistance were harvested in this research that facilitate the understanding of Bt toxin functions in plant transgenic breeding.
Bt Cry1Ac22 toxin; Bt toxin functional domain; Plant expression construct; Arabidopsis thaliana; Genetic transformation