摘要
目的 :构建颗粒溶解素 (granulysin)cDNA原核表达载体并进行表达。 方法 :采用逆转录聚合酶链反应 (RT PCR)技术获得颗粒溶解素cDNA ,克隆到pGEM T载体 ,测序正确后 ,再亚克隆到质粒 pGEX 4T1中 ,构建重组表达载体 pGEX 4T1 granulysin ,并转化大肠埃希菌DH5α,诱导GST granulysin融合蛋白表达。 结果 :诱导含重组表达载体pGEX 4T1 granulysin的大肠埃希菌DH5α后 ,免疫印迹显示出一条相对分子量 (Mr)为 4 4 0 0 0的蛋白条带。结论 :获得了颗粒溶解素与GST的融合蛋白 。
Objective:To construct prokaryotic expression vector carrying granulysin cDNA and to express it in E.coli. Methods:Using RT PCR technique,ganulysin cDNA was obtained,cloned into clone vector pGEM T,sequenced and then cloned into expression vector pGEX 4T1.Thus the prokaryotic expression vector pGEX 4T1 granulysin was constructed,and E.coli DH5α was transformed,induced to obtain a fusion protein.Results:After inducing E.coli DH5α carrying expression vector pGEX 4T1 granulysin,it was showed that a fusion protein with relative molecular mass (Mr) 44 000 was detected by Western blot analysis.Conclusions:A granulysin GST fusion protein is obtained,so as to provide an experiment basis for futher research.
出处
《蚌埠医学院学报》
CAS
2003年第4期283-285,共3页
Journal of Bengbu Medical College
基金
安徽省教育厅自然科学研究重点项目 (No :99J10 15 0zd)
