摘要
采用PCR技术 ,删除了小鼠CREBcDNA5′端和 3′端非编码序列 ,并引入便于基因操作的酶切位点。经 30次循环的扩增 ,得到改造后的CREBcDNA ,全长 10 71bp。亚克隆后 ,对此扩增片段进行了限制性内切酶物理图谱分析 ,测定了DNA序列 ,并以其为插入物 ,构建了 pBV2 2 0 -PCR -CREB重组表达载体。经Western印迹法分析证明 。
The mouse CREB cDNA was reformed by PCR technique to delete 5'\|and 3'\|terminal nontranslating regions affecting its expression and to introduce restriction sites for genetic manipulation.The CREB cDNA(1071bp)reformed was abtained after 30 amplification cycles and was analyzed for its restriction map and its sequence.Expression vetors of the cDNA in E.coli were constructed.the two recombinant vectors were successfully expressed in E.coli on the basis of showing by western blotting.\;
出处
《生物工程进展》
CSCD
2000年第2期55-57,共3页
Progress in Biotechnology
