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从大豆 (花生豆 1号 )未成熟种子中提取总RNA ,逆转录形成cDNA第一条链。根据genebank中大豆 11S球蛋白Gy5的cDNA序列设计引物 ,用PCR方法扩增Gy5的cDNA序列 ,克隆到 pUC19质粒载体上 ,并测定其全序列。用限制性内切酶PstI和EcoRI酶切重组质粒 ,将目的片段与质粒 pCAMBIA130 1连接 ,构建成植物表述载体并转化到农杆菌中 ,以便于以后的研究利用。
Total RNA was extracted from immature seeds of soybean (peanut beans 1) and reverse transcribed to form the first strand of cDNA. Primers were designed according to the cDNA sequence of soybean 11S globulin Gy5 in genebank. The cDNA sequence of Gy5 was amplified by PCR and cloned into pUC19 plasmid vector. The full sequence of Gy5 was obtained. The recombinant plasmids were digested with restriction enzymes PstI and EcoRI. The target fragment was ligated with plasmid pCAMBIA130 1 to construct plant expression vector and transformed into Agrobacterium for further study.