摘要
目的:构建人纤溶酶真核表达载体以用于抗栓、溶栓的研究。方法:人胎肝组织经PCR技术扩增人纤溶酶基因,定向克隆至真核表达质粒pC-DNA-3后测序。经测序鉴定正确后构建人纤溶酶真核表达重组质粒,利用PCR和酶切方法鉴定重组真核表达质粒的正确性。结果:目的基因PCR扩增产物为1 740 bp,测序结果显示,目的基因PCR扩增产物与GenBank登记的序列完全相同。PCR和酶切鉴定结果显示,构建的重组真核表达质粒中含有正确编码的人纤溶酶基因全长序列。结论:成功构建了含有人纤溶酶基因的真核表达重组质粒,并进行了鉴定。
Objective To construct the eukaryotic expression vector of human fibrinolysin for further study on antithrombus and thrombolysis. Methods Human fibrinolysin gene was amplified from human fetal liver tissue with PCR, and was cloned into eukaryotic expression plasmid pCI-neo and then sequenced. After the correct identification by sequencing, the eukaryotic expression recombinant plasmid of human fibrinolysin was constructed and identified with PCR and enzyme digestion. Results The PCR amplification product of fibrinolysin gene was 1 740 bp. The result of sequencing was the same as that registered in GenBank. The results of PCR and enzyme digestion showed that the recombinant eukaryotic expression plasmid had correct codogenic gene fragment. Conclusion The eukaryotic expression recombinant plasmid of human fibrinolysin gene is successfully constructed and identified.
出处
《吉林大学学报（医学版）》
CAS
CSCD
北大核心
2009年第5期898-901,共4页
Journal of Jilin University：Medicine Edition
基金
长春市科技计划项目资助课题(2006-2009)
关键词
人纤溶酶
抗栓
真核表达载体
重组质粒
human fibrinolysin
anti-thrombus
eukaryotic expression vector
recombinant plasmid
