大豆胰蛋白酶抑制剂KSTI3基因的克隆及其植物表达载体的构建  

Cloning and Construction of Expression Vector on KSTI3 Gene of Soybean Kunitz-type Trypsin Inhibitor

作　　者：吕品[1] 柴晓杰[2] 王丕武[1] 张宇[1] 

机构地区：[1]吉林农业大学生物技术学院,长春130118 [2]大连水产学院生命科学与技术学院,大连116023

出　　处：《吉林农业大学学报》2007年第3期275-278,共4页Journal of Jilin Agricultural University

基　　金：吉林省重大科技攻关项目(20040203-2-2)

摘　　要：以大豆基因组DNA为模板,利用聚合酶链式反应(PCR)技术克隆了大豆胰蛋白酶抑制剂基因KSTI3的全长DNA片段,并将其构建到pMD18-T vector上。核苷酸序列测定结果表明:该基因片段全长654 bp,与已发表的KSTI3基因序列同源性达99%。将反义+正义基因片段插入到pBI121 35S启动子下,构建重组质粒pBIKSTI3。通过冻融法将该重组质粒转入根癌农杆菌EHA105中,获得了植物表达载体。Soybean Kunitz-type trypsin inhibitors are important anti-nutritious factors. The full-length DNA fragment of soybean Kunitz-type Trypsin Inhibitor gene KSTI3 was amplified by PCR technique, then PCR products were cloned into pMD18-T vector. Nucleotide sequence analysis showed that the cloned fragment consisted of 654 bp and shared 99% identity with the reported KSTI3. The fragment of anti-sense gene ＋ plus-sense gene was constructed to the downstream of 35 S promoter in the binary vector pBI121, which formed the recombinant plasmid pBIKSTI3. The plasmid pBIKSTI3 was mobilized into Agrobacterium tumefaciens strain EHA105 by freeze-melting methods, and a plant expression vector was thereby obtained.

关 键 词：胰蛋白酶抑制剂 PCR 克隆 植物表达载体 大豆 

分 类 号：Q785[生物学—分子生物学]