摘要
目的:构建含人端粒酶逆转录酶(Humantelomerasereversetranscriptase,hTERT)基因不同位点的一系列siRNA真核表达质粒mU6-hTERT1～5,为以端粒酶为靶点的基因治疗鼻咽癌研究奠定基础。方法:将人工合成的针对hTERT基因4个不同位点和1个混乱序列的正反链DNA序列经基因重组定向克隆插入到真核表达质粒载体mU6中,通过PCR检测确定重组结果。根据260nm处紫外红吸收值计算重组质粒的浓度。结果:各对DNA序列均按正确方向插入pmU6质粒,纯度和浓度均符合实验要求。结论:成功构建成了含人端粒酶hTERT基因4个不同位点的siRNA真核表达质粒。
Objective： To construct a series of the eukaryotic expression plasmid mU6-hTERT1 to mU6-hTERT5 which can transcript siRNA against human telomerase＇s hTERT gene for future study of the Nasopharyngeal neoplasm＇s gene therapy. Methods： 5 pairs of sense and antisense sequences of siRNA that 4 pairs were against different sites of hTERT gene and 1 pair was scrabble DNA sequences were synthesized. They were inserted into the eukaryotic expression vector pmU6 with definite direction. Their purity and density were done and determined by mesuring the absorbance at 260 nm and 280 nm. Results： The siRNA sequences were successfully inserted into the eukaryotic expression vector pmU6. The reeombinated plasmids were identified by PCR test and DNA sequence. The puprity and concentration of the series of plasmids were inspected. Conclusion： 5 siRNA transeripting plasmids pmU6-hTERT1 to pmu6-hTERT5 are successfully constructed.
出处
《海南医学院学报》
CAS
2005年第4期266-268,共3页
Journal of Hainan Medical University
基金
广东省自然科学基金资助项目(NO.31962)
