VEGFR-3第二免疫球蛋白样区(2-Ig)反义多肽的基因克隆与原核表达  

Cloning and Prokaryotic Expression of the Antisense Peptide of VEGFR-3 2-Ig

作　　者：李书华[1] 张雅洁[1] 李冰[2] 张惠球[1] 李杰荣[1] 

机构地区：[1]广州医学院病理学教研室,广东广州510182 [2]广州医学院中心实验室,广东广州510182

出　　处：《广州医学院学报》2003年第3期23-28,共6页Academic Journal of Guangzhou Medical College

基　　金：广州市教育局科研基金资助

摘　　要：目的 :构建 pGEX 6P 3 VEGFR 3原核表达载体 ,获得纯化的VEGFR 3胞外第二免疫球蛋白样区 (2 Ig)反义多肽。方法 :通过RT PCR技术从 1 6周胎龄胚胎肺组织中扩增VEGFR 32 IgcDNA ,反向克隆至 pGEX 6P 3载体 ,用IPTG诱导重组菌株表达 ,用Sepharose 4B亲和层析柱对表达产物进行纯化 ,PreScissionProtease酶对融合蛋白进行解离。结果 :构建 pGEX 6P 3 VEGFR 3原核表达载体 ;分离纯化出约 1 2kD的VEGFR 32 Ig反义多肽。结论 :成功获得VEGFR 3胞外第二免疫球蛋白样区 (2 Ig)反义多肽 ,为研究VEGFR 3在肿瘤生长和转移中的作用奠定基础。Objective:To construct the prokaryotic expression plasmid and acquire the antisense peptide of VEGFR 3 2 Ig. Methods:Amplify the gene of VEGFR 3 2 Ig from the 16w embryo lung through the RT PCR,then clone its antisense into the pGEX 6P 3 plasmid .The recombinant vector was induced by the IPTG and the expression product was purified through the Sepharose 4B column. The VEGFR 3 2 Ig antisense peptide can be cut from the fusion by PreScission Protease. Results:Construction of the recombinant plasmid pGEX 6P 3 VEGFR 3 and acquisition of the VEGFR 3 2 Ig antisense peptide were finished sucessfully. Conclusion:Successful acquisition of VEGFR 3 2 Ig antisense peptide would be useful in the studying the role of VEGFR 3 in cancer growth and metastasis.

关 键 词：VEGFR-3 2-Ig 反义多肽 原核表达 

分 类 号：R392[医药卫生—免疫学]