人胰岛素基因的克隆及其乳腺特异性表达载体的构建

Cloning and Vector Construction of Human Insulin Gene

从人血中提取人基因组DNA,根据GenBank登陆的人胰岛素基因序列设计引物,通过longPCR技术克隆了人胰岛素基因,并将该基因克隆入pMD18T载体构建pMI,将pMI和质粒pBEBT通过酶切连接构建成重组质粒pBEBI。结果成功扩出长为1.6kb的人胰岛素基因并构建了乳腺特异性表达载体。

To clone human insulin gene and construct mammary expressing vector the human insulin genomic DNA fragment was amplifed from human genomic DNA with long PCR technolog.We constructed pMI by inserting the insulin gene into the T site of pMD18T veetor.pMI and pBEBI were digested respectivly by restriction endonuclease Nhel and BanH I.Then,the two fragments were harvested and ligated to construct the recombinant vector.