【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。更多还原

【Abstract】 Soybean Kunitz-type trypsin inhibitors are important antinutritious 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.更多还原