【摘要】 目的:克隆人TGFBI基因,并构建其真核表达载体。方法:通过RT-PCR从供体角膜移植术后留下的角膜组织中克隆人TGFBI基因全长编码区,将该DNA片断亚克隆到克隆载体pMD18-T中,进一步通过和SalI双酶切将目的片断定向克隆至真核表达质粒载pCI中。经PCR,酶切和序列测定方法鉴定重组质粒。结果:获得了人TGFBI的编码基因,并成功建了其真核表达载体。结论:人TGFBI基真核表达质粒的成功构建并鉴定。更多还原

【Abstract】 AIM:To clone human TGFBI gene and construct its eukaryotic expression plasmid. METHODS:The full coding domain sequence of TGFBI gene was cloned from human corneal tissue from operative spaceman with reverse transcriptase-polymerase chain reaction (RT-PCR) and was sub-cloned into pMD18-T. The targeted DNA fragment,digested with restriction enzyme XbaI and SalI,was directionally cloned into eukaryotic expression plasmid pCI. Then the reconstructed plasmid was identified with PCR,enzyme digestion and sequencing. RESULTS:Human TGFBI gene was successfully amplified and its eukaryotic expression plasmid was constructed. CONCLUSION:The successive reconstruction and verifying of eukaryotic expressive plasmid containing human TGFBI gene established the foundation of studying of the mechanisms of TGFBI gene mutation related corneal dystrophy.更多还原