人DcR3cDNA克隆及其在COS-7细胞中的表达  

Cloning and expression of human DcR3 molecule in COS-7 cells

作　　者：王槐志[1] 黎万玲[2] 董家鸿[1] 倪兵[2] 姜曼[2] 吴玉章[2] 

机构地区：[1]第三军医大学西南医院全军肝胆外科研究所 [2]第三军医大学基础医学部全军免疫学研究所,重庆400038

出　　处：《第三军医大学学报》2006年第17期1749-1751,共3页Journal of Third Military Medical University

基　　金：国家自然科学基金资助项目(30400418);重庆市自然科学基金资助重点项目(CSTC;2005BA5004)

摘　　要：目的克隆人DcR3分子的cDNA,将其重组入真核表达载体,并表达于COS-7细胞。方法以PCR方法克隆编码人DcR3分子的cDNA,对其序列进行测定。将测序正确的DcR3cDNA克隆入真核表达载体pAAV-IRES-hrGFP,构建重组表达载体。采用脂质体法转染COS-7细胞,用Western blot检测和激光共聚焦检测重组DcR3分子在COS-7细胞中的表达。结果PCR方法扩增出一1000bp左右的基因片段,插入pAAV-IRES-hrGFP构建重组表达载体后,经序列测定证实扩增的片段为人DcR3分子cDNA。将重组子转染COS-7细胞,Western blotting和激光共聚焦检测到DcR3分子的表达。结论成功地克隆并在COS-7细胞中表达了DcR3分子。Objective To clone ORF of DcR3 gene and insert it into eukaryotic expression vector and express it in COS-7 cells. Methods Encoding sequence of human DcR3 gene was cloned by PCR and sequenced. The sequenced ORF was cloned into eukaryotic expression vector pAAV-IRES-hrGFP to construct recombinant plasmid. COS-7 cells were transfected with recombinant plasmid by lipofectamine2000. Expression of recombinant DcR3 gene was verified by Western blotting and confocal microscopy. Results A 1 000-bp gene segment was obtained by PCR and inserted into pAAV-IRES-hrGFP segment was proved to be encoding sequence of human DcR3 gene cells was verified by Western blotting and confocal microscopy. cloned and expressed in COS-7 cells.

关 键 词：DCR3 COS-7细胞 基因表达 

分 类 号：R392.11[医药卫生—免疫学]