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Expression of the Spinach Betaine Aldehyde Dehydrogenase (BADH) Gene in Transgenic Tobacco Plants

Liang Zheng 1\ Ma Deqin 2\ Tang Lan 1\ Hong Yiguo 2\ Luo Ailing 1\ Dai Xiuyu 2 (Institute of Botany,Academia Sinica,Beijing 100093) 1 (Institute of Microbiology,Academia Sinica,Beijing 100080) 2  
Plasmid pLS9 contains a 1.5 kb of spinach BADH cDNA including its complete open reading frame.The 1.5 kb of BADH cDNA was cutted from pLS9 using restriction enzyme and was inserted into the expression cassette of pYH between the 35S promoter and polyA terminator.The 35S BADH gene polyA fragment was cloned into binary vector pBin19 in the polylinker site.Resulting plasmid pBinBADH S was introduced into Agrobacterium tumefaciens LBA4404.The BADH gene was transformed to tobacco plants mediated by agrobacteria.Ninety kanamycin resistant transformants were selected.PCR detection showed that more than 60% of transformed tobacco plants contain the foreign BADH gene.Western blot analysis ,BADH activity assay and BADH specific activity stain showed that BADH gene was normally expression in the transgenic tobacco plants.The enzyme also occured in chloroplasts and the cytosol in the transgenic plants.Plants having strong expression of BADH gene had an ability to tolerate high salinity.
【Fund】: 国家自然科学基金
【CateGory Index】: Q81
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