NOS/HuIFNa嵌合基因的构建及其与Ti质粒载体的拼接

CONSTRUCTION OF NOS/HuIFNa CHIMERIC GENE AND ITS FUSION WITH Ti PLASMID VECTOR

将人α干扰素(HuIFNa)基因的全长cDNA克隆到nos启动子的下游,并在3＇端加上nos的3＇末端片段,得到人α干扰素基因插入方向同nos启动子一致的重组质粒pLQP24和方向相反的重组质粒pLQN2,并分别同Ti栽体pARCl2拼接。将NOS/HuIFNa引入到pARCl2的T区端臂内,构建成重组质粒pLPP3和pLNP20通过接合转移将这两个重组质粒引入到根癌土壤杆菌LBA4404。

The lull length cDNA of HuIFNa gene was inserted between the downstream of nos promoter and the upstream of 3' end. The two recombinant plasmids with opposite orientation were obtained, one was named as pLQP24, which carries the chimerie gcne: nos promoter——HuIFNa gone(5'-3')-nos 3' end, another was named as pLQN2, which carries the chimeric gent: nos promoter-HuIFNa(3' -5')-nos 3' end. The pLQP24 and pLQN2 was fused separately with pARC12 by the unique HindⅢ site on them. The two recombinant plasmids, pLPP3 and pLNP20, were obtained and conjugated respectively into Agrobacterium tumefaciens LBA4404.