人核心蛋白聚糖真核表达载体的构建及鉴定  

Construction and identification of human decorin gene recombinant eukaryotic expressing vector

作　　者：操海萍[1] 舒振波[2] 张桂珍[1] 

机构地区：[1]吉林大学中日联谊医院中心实验室 [2]吉林大学中日联谊医院普通外科,吉林长春130033

出　　处：《吉林大学学报（医学版）》2006年第4期590-592,共3页Journal of Jilin University：Medicine Edition

基　　金：吉林省科技厅科技发展计划资助课题(20031268)

摘　　要：目的:构建人核心蛋白聚糖(DCN)真核表达载体,为进一步研究其生物学活性奠定基础。方法:采用聚合酶链反应(PCR)扩增目的片段,PCR产物及真核表达载体pcDNA3分别双酶切后进行连接,并转化入大肠杆菌JM109中扩增以获得重组载体。结果:PCR获得与预期大小一致的、约1 000 bp的特异性DNA片段,PCR产物与表达载体经双酶切后连接,构建重组载体pcDNA-dec,经双酶切鉴定及测序证实,人DCN基因cDNA片段正确插入真核表达载体中。结论:成功构建人DCN真核表达载体pcDNA-dec。Objective To construct a recombinant eukaryotic expressing vector pcDNA-dec and provide a basis for further study on the bioactivity of decorin （DCN）. Methods DCN cDNA was amplified by using polymerase chain reaction （PCR）. Product of PCR and expressing vector were digested by restriction endonucleases, then ligated and transformed into JM109 bacteria. Results The specific DNA fragment was obtained by PCR as supposed. Product of PCR and expressing vector were digested by restriction endonucleases, then ligated to establish the recombinant eukaryotic expression vector pcDNA-dec. It was confirmed that DCN cDNA was inserted into the eukaryotie expression vector correctly by using digestion identification and sequencing. Conclusion The recombinant eukaryotic expression vector pcDNA-dec of human DCN is successfully constructed.

关 键 词：核心蛋白聚糖 基因表达 聚合酶链反应/方法 

分 类 号：Q786[生物学—分子生物学]