HD-5分泌性表达载体构建  

Construction of secretary expression vector of HD-5

作　　者：谭虎[1] 粟永萍[2] 杨天德[1] 艾国平[2] 黄岚[3] 陶军[1] 

机构地区：[1]第三军医大学新桥医院麻醉科,重庆400037 [2]第三军医大学预防医学系全军复合伤研究所,创伤、烧伤与复合伤国家重点实验室,重庆400038 [3]第三军医大学新桥医院心内科,重庆400037

出　　处：《重庆医学》2007年第16期1585-1586,共2页Chongqing medicine

基　　金：国家自然科学基金重点资助课题(30230360);创伤烧伤复合伤国家重点实验室开放资助课题(30470527)

摘　　要：目的构建防御素5分泌性表达载体。方法通过PCR重叠延伸技术得到HD-5信号肽及成熟肽的嵌合分子,并将其插入真核表达质粒pVITRO3。结果测序结果显示,成功拼接HD-5的信号肽及成熟肽,并插入到真核表达质粒pVITRO3,阅读框正确。结论PCR重叠延伸技术适用于分泌性载体构建。Objective To construct the secretary expression vector of mutation chimeric of human defensin 5 （HD-5） by clone technology. Methods The PCR product of signal and mature peptides of HD-5 were linked by splicing by overlap extension（SOE）. Then the chimeric HD-5 was cloned into pVITRO3 vector. The constructed vector pVITRO3-HD-5 was identified by restriction endonucleases and sequenced. Results The result of the suquencing showed the sequence of signal peptide and muture peptide of HD-5 was connected successfully and was cloned into eukaryotic plasmid. Conclusion SOE is appropriate for establishing secretion type of expression vector.

关 键 词：防御素-5 基因克隆 PCR重叠延伸技术 肠源性感染 

分 类 号：Q78[生物学—分子生物学]