论文部分内容阅读
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 .
OBJECTIVE: To construct a hairpin ribozyme gene against human vascularendothelial growth factor (VEGF) and to detect its cleavage activity in vitro.Methods Human VEGF ribozyme gene was cloned into the eukaryotic expression vector pcDNA3 , And simultaneously in vitro transcribed pcDNA3-RZ, detection of ribozyme cleavage function.Results Eco RI and Bam H digestion confirmed that ribozyme gene fragments of the correct size, the resulting ribozyme cutting activity.Conclusion successfully constructed Anti-human VEGF hairpin ribozyme gene eukaryotic expression vector, laid the foundation for the next gene therapy.