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玉米特异启动子驱动下结核Hsp65与Esat-6融合基因表达载体的构建及鉴定

DOI: 10.7668/hbnxb.2009.03.006, PP. 28-31

Keywords: 结核分支杆菌,热休克蛋白65,Esat-6,globulin-1

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Abstract:

利用转基因玉米研制新型结核口服疫苗.构建植物双元表达质粒pCAMBIA1300GHLE,并转化农杆菌LBA0.以本实验室构建的pEGHLE为模板经聚合酶链反应(PCR)扩增出Hsp65-Esat6基因,连接到含有玉米特异性启动子globulin-1的pCR2.1载体上;将globulin1-HLE联合片断切下连到含有抗除草剂基因bar的pCAMBIA1300载体中;电击法将重组质粒转化到农杆菌LBA0中.成功构建了pC1300GHLE质粒,酶切鉴定得到3.3kb和8.6Kb2条带,测序分析表明克隆的H8p65和Esat6序列与NCBI上公布序列一致;成功转化到农杆菌中,酶切从农杆菌中所提的质粒,条带大小与预期结果相符合.成功构建和转化了pCl300GHLE表达载体,为成功研制利用转基因植物生产抗结核口服疫苗奠定了基础.

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