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中文摘要：
目的:构建人CyPA基因重组质粒pET30α(+)-CyPA。方法:根据CyPA基因设计引物,从人肺癌组织cDNA中扩增目的基因。pET30α(+)质粒、目的基因DNA经NdeI酶、XhoI酶双酶切,用DNA快速连接试剂盒连接得重组子pET30α(+)-CyPA。将重组子转化DH5α,经卡那霉素琼脂糖平板筛选,挑选阳性克隆进行PCR扩增并测序。结果与结论:快速PCR能从阳性克隆中扩增出目的DNA。将重组质粒测序,由起始密码ATG到终止密码TAG止,扩增得到的目的基因片段具有完整的阅读框;与人CyPA基因cDNA序列进行比较,同源性达到99.8%。重组质粒pET30α(+)-CyPA构建成功。
英文摘要：
Aim:To construct the recombinant plasmid of pET30α(+).Methods:The primer was designed according to CyPA gene sequence,CyPA gene was amplified from cDNA of human lung cancer tissue. The CyPA gene and the pET30α(+) were digested by Nde I,Xho I,and the CyPA gene fragments and the plasmids of pET30α(+) were ligated with T4 DNA rapid ligase to form the recombinants pET30α(+)-CyPA,then the products were tansfered into the competent DH5α. The transformants were screened by kanamycin and were identified by rapid PC...
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