【摘要】 根据已知序列(GeneBankX076075),设计引物,用PCR方法获得了一段大豆叶绿体DNA片段,命名为soy。将来源于质粒pBluescriptSK( +)的含氨苄抗性基因Ampr和大肠杆菌质粒复制起始点ColE1ori的一段DNA片段克隆到这段大豆叶绿体DNA片段中,然后与由三个基因(壮观霉素抗性基因aadA、甘露聚糖酶基因man及绿色荧光蛋白基因gfp)串联的表达盒相连,以构建大豆叶绿体多顺反子表达载体pCS。并在大肠杆菌中通过平板定性分析和Western印迹的方法对所构建载体上的表达盒进行了功能鉴定,结果表明同一多顺反子的三个基因均得到了表达。更多还原

【Abstract】 A soybean choloroplast DNA fragment named soy was isolated by PCR based on the published sequence (GeneBank X076075) and inserted into a fragment containing Amp~r and ColE1 ori from pBluescript SK(+) to generate soybean chloroplast expression vector pCS with an expression cassette containing three tandem genes (aadA, man and gfp). The function of the expression cassette was identified by plate qualitative assay and Western blotting in E.coli. The results showed that the multicistrons were expressed effectively through soybean chloroplast expression vector pCS.更多还原