Pr1基因和绿色荧光蛋白基因融合表达载体的构建

摘要

Objective:To construct the expression vector contained fusion gene Pr1 and enhanced green fluorescent protein (EGFP).EGFP is the reporter gene,and Pr1 is the target gene.Methods:The Pr1 was connected to the upstream of EGFP and the downstream of EGFP respectively.The fusion gene of EGFPPr1 and Pr1-EGFP was constructed,and then the fusion gene was inserted to the vector pTWIN1 respectively.Results:Induced by IPTG,the recombinant showed brightly green fluorescent when excited by long-wave ultraviolet light,the Pr1 can be detected from the induced strains after crushing.Conclusion:The expression vector was constructed successfully through genetic engineer technique.It is useful to research on Pr1 by EGFP using as biology marker molecular.%目的:以增强型绿色荧光蛋白(Enhanced Green Fluorescent Protein,EGFP)基因为报告基因,类枯草菌素蛋白酶(subtilisin-like protease,Pr1)基因为目的基因,分别构建含有EGFP-Pr1和Pr1-EGFP融合基因的原核表达载体.方法:将Pr1基因分别连接到EGFP基因的上游和下游,插入质粒pTWIN1,构建表达载体pEGFP-Pr1和pPr1-EGFP.结果:经异丙基硫代半乳糖苷(IPTG)诱导后,重组菌株在长波紫外线的照射下,均能发出明亮的绿色荧光,并且诱导菌株破碎后均可检测到Pr1蛋白酶的活性.结论:应用基因工程技术成功构建pPr1-EGFP和pEGFP-Pr1融合基因表达载体,EGFP作为Pr1生物标记分子,有利于进一步开展Pr1基因的特异性功能研究.