CAJ | 학술논문

目的:构建pcDNA3.1/Cx50 V64G真核表达载体,用生物信息学软件分析V64G突变对人缝隙连接蛋白Cx50的影响.方法:经PCR获得Cx50基因片段,重组至真核表达载体pcDNA3.1/Amp(+)中,经PCR,酶切和序列测定方法鉴定重组质粒.结果:获得了具有V64G突变的Cx50的编码基因,并成功构建了其真核表达载体.并证明64位缬氨酸在不同物种的Cx50及在人的多种缝隙连接蛋白是高度保守的区域,与白内障的发生高度相关.结论:pcDNA3.1/Cx50 V64G真核表达质粒的成功构建和鉴定为进一步研究白内障机理奠定了基础.
목적:구건pcDNA3.1/Cx50 V64G진핵표체재체,용생물신식학연건분석V64G돌변대인봉극련접단백Cx50적영향.방법:경PCR획득Cx50기인편단,중조지진핵표체재체pcDNA3.1/Amp(+)중,경PCR,매절화서렬측정방법감정중조질립.결과:획득료구유V64G돌변적Cx50적편마기인,병성공구건료기진핵표체재체.병증명64위힐안산재불동물충적Cx50급재인적다충봉극련접단백시고도보수적구역,여백내장적발생고도상관.결론:pcDNA3.1/Cx50 V64G진핵표체질립적성공구건화감정위진일보연구백내장궤리전정료기출.
AIM : To construct and analyze eukaryotic expression plasmid inserted by Cx50 with V64G mutation through bioinformatics software.METHODS: The full coding domain sequence of Cx50 with V64G mutation was acquired from the blood of patients with cataract and was cloned into pcDNA3.1 /Amp (+).The constructed plasmid was identified with PCR , enzyme digestion and sequencing. The analysis of Cx50 with V64G mutation was performed with bioinformatics software.RESULTS : Cx50 with V64G mutation was successfully amplified and its eukaryotic expression plasmid was constructed. Valine-64 is well conserved in the first extracellular loop of connexin 50 in different species and also in different human α -type gap junctional proteins.CONCLUSION : The successive reconstruction and verification of eukaryotic expression plasmid containing Cx50 with V64G mutation established the foundation for further studying the mechanism of cataract.