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.
出处
《第四军医大学学报》
CAS
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)
关键词
肿瘤
基因治疗
血管内皮生长因子
核酶
gene therapy
vascular endothelial growth factor
ribozyme
