抗人VEGF发夹状核酶基因的合成、克隆、表达及活性鉴定  

Synthesizing, cloning, expressing of anti-vascular endothelial growth factor hairpin ribozyme gene and assaying of its activity

作　　者：金明[1] 严瑞兰[2] 赵君庸 惠宏襄[1] 王成济[1] 杨安钢[1] 

机构地区：[1]第四军医大学基础部生物化学与分子生物学教研室,陕西西安710033 [2]第四军医大学西京医院妇产科,陕西西安710033 [3]西安交通大学医学院生物化学教研室,陕西西安710061

出　　处：《第四军医大学学报》2000年第11期1421-1423,共3页Journal of the Fourth Military Medical University

基　　金：国家自然科学基金!资助项目 (3970 0 14 7)

摘　　要：目的　构建抗人血管内皮生长因子 (vascularendothelial growth factor,VEGF)发夹状核酶基因 ,并在体外检测其切割活性 .方法　化学法合成的人 VEGF核酶基因定向克隆入真核表达载体 pc DNA3中 ,并在体外同时转录pc DNA3- RZ,检测核酶的切割功能 .结果　经 Eco RI和Bam H 酶切鉴定证实核酶基因片段大小正确 ,所产生的核酶具有切割活性 .结论　成功地构建了抗人 VEGF发夹状核酶基因真核表达载体 ,为下一步进行基因治疗打下了基础 .AIM To construct expression vector bearing anti vascular endothelial growth factor (VEGF) hairpin ribozyme (RZ) gene and to assay the cleavage activity of the ribozyme in vitro . METHODS Anti VEGF hairpin RZ gene was synthesized and inserted to the eukar yotic expression vector pcDNA3. Then pcDNA3 RZ was transcripted in vitro and the cleavage activity of the RZ was measured. RESULTS Fregment of the RZ gene was confirimed by the digestion of pcDNA3 RZ with Eco RI and Bam HI. The expression product the RZ gene had cleavage activity. CONCLUSION We have successfully constructed pcDNA3 RZ expression vector and laid a basis for further study of gene therapy.

关 键 词：肿瘤 基因治疗 血管内皮生长因子 核酶 

分 类 号：R730.59[医药卫生—肿瘤]