菠菜甜菜碱醛脱氢酶基因在烟草中的表达  

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

作　　者：梁峥[1,2] 马德钦[1,2] 汤岚[1,2] 洪益国 骆爱玲 戴秀玉[1,2] 

机构地区：[1]中国科学院植物研究所 [2]中国科学院微生物研究所

出　　处：《生物工程学报》1997年第3期236-240,共5页Chinese Journal of Biotechnology

基　　金：国家自然科学基金

摘　　要：质粒ｐＬＳ９含有１５ｋｂ的编码菠菜甜菜碱醛脱氢酶（ＢＡＤＨ）基因。经限制酶切后克隆到植物表达载体的３５Ｓ启动子和ＰｏｌｙＡ终止子之间。经农杆菌介导转化烟草，获得９０多株抗卡那霉素再生植株，经ＰＣＲ检测证明６０％以上再生植株含有ＢＡＤＨ基因。转基因植株经Ｗｅｓｔｅｒｎｂｌｏｔ，ＢＡＤＨ酶活性测定，ＢＡＤＨ酶活性特异性染色法检查和耐盐性分析，证明菠菜ＢＡＤＨ基因在烟草正常表达，在叶绿体和胞液中均有ＢＡＤＨ酶存在。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.

关 键 词：烟草 甜菜碱醛脱氢酶 基因表达 转基因烟草 

分 类 号：S572.032[农业科学—烟草工业] S188[农业科学—作物学]