【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SKTI3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SKTI3基因序列同源性达99%。将目的基因片段插入到pB I121 35S启动子下,构建重组质粒pB ISKTI3,采用冻融法将该重组质粒转入农杆菌EHA105中,获得了植物表达载体。更多还原

【Abstract】 The gene SKTI3 of soybean Kunitz-type trypsin inhibitor was cloned by PCR using soybean genomic DNA as template and inserted into pMD18-T vector.DNA sequence analysis showed that the cloned fragment consisted of 654 bp and shared 99% identity with the SKTI3 sequence published on Genbank.The cloned gene was inserted to the downstream of 35S promoter in the binary vector pBI121 to construct the recombinant plasmid pBISKTI3.Then,pBISKTI3 was mobilized into Agrobacterium tumefaciens strain EHA105 by freeze-melting methods.The plant expression vector containing SKTI3 gene was obtained.更多还原