人源性高血糖素原基因的克隆  

cDNA cloning of human proglucagon from brain tissue

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400038

出　　处：《重庆医学》2007年第16期1580-1581,共2页Chongqing medicine

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

摘　　要：目的克隆人高血糖素原基因(proglucagonc DNA,PG cDNA),构建其真核表达质粒载体。方法从人切除脑组织中提取总RNA,并分离mRNA,经过逆转录-聚合酶链反应(RT-PCR)扩增出PGcDNA,并将其插入经BamHⅠ和EcoRⅠ双酶切的真核表达质粒载体pVITRO3中,构建重组质粒载体pVITRO3-PGcDNA。结果经过限制性酶切鉴定和测序证明,PGcD-NA全长543bp,编码181个氨基酸,并已插入到pVITRO3中。结论成功获得了PGcDNA,构建了含PGcDNA的真核表达质粒载体,为研究重组PG的功能奠定了基础。Objective To construct the recombinant eukaryotic plasmid of human proglucagon （PG） and to prepare for expressing in eukaryotic cells. Methods Total RNA as well as mRNA were isolated from human brain tissue and eDNA encoding PG was amplified by reverse transcription PCR （RT-PCR）. The eDNA was inserted into the vector pVITRO3. The constructed vector pVITRO3-PG eDNA was identified by restriction endonucleases and sequenced. Results No discrepancy was found in nucleotide as compared with standard sequence of PG from GenBank. It encoded 229 amino acids, whose length was 687 bp. Conclusion The eDNA of PG is successfully obtained and the recombinant plasmid is constructed correctly. It will be helpful for analyzing the function of recombinant PG.

关 键 词：高血糖素原 聚合酶链式反应 基因表达 

分 类 号：Q78[生物学—分子生物学]