Construction of a plant expression vector of the CryⅢA gene and its genetic transformation in potato

doi:10.11686/cyxb20140130

Abstract

Abstract: Using In-Fusion technology,plant expression vectors were constructed from the vector pBI121 for transformation of the CryⅢA gene under the control of the constitutive promoter CaMV 35S or the potato leaf and stem-specific promoter ST-LS1. The recombinant plasmids were introduced into Agrobacterium tumefaciens strain LBA4404 by the freeze-thaw method. The putative transgenic plants of potato cultivars Longshu No.3 and Gannongshu No.2 were obtained by the Agrobacterium-mediated transformation system. Results from selection of the transformed plants on culture media containing kanamycin and PCR assay showed that the CryⅢA gene was integrated into the genome of 10 potato plants. Real-time fluorescence quantitative PCR (qRT-PCR) analysis indicated that the CryⅢA gene driven by the promoter CaMV 35S,was expressed in roots,stems and leaves,with a high expression in leaves of the transgenic plants. The CryⅢA gene driven by the promoter ST-LS1 was expressed only in stems and leaves with the highest expression in leaves of the transgenic potato plants. However,no traces of it were found in roots of the transgenic potato plants.

CLC Number:

S435.32

YOU Jia,ZHANG Ning,WEN Yi-kai,WU Jia-he,SI Huai-jun,WANG Di. Construction of a plant expression vector of the CryⅢA gene and its genetic transformation in potato[J]. Acta Prataculturae Sinica, 2014, 23(1): 248-256.

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