分子生物学
融合蛋白
重组DNA
免疫印迹
生物
轮状病毒
质粒
表达式向量
病毒学
拉伤
基因
大肠杆菌
载体(分子生物学)
病毒
生物化学
解剖
作者
Nan Guo,Xiaoman Sun,Dandi Li,Youde Cao
出处
期刊:Chinese Journal of Clinical Hepatology
日期:2015-10-30
卷期号:29 (5): 452-454
标识
DOI:10.3760/cma.j.issn.1003-9279.2015.05.016
摘要
Objective
To express the VP8* protein of the rotavirus vaccine strain LLR in the E. coli with the pGEX4T-1 vector, which is important for the further research of the VP8* protein functions.
Methods
The LLR VP8* gene was obtained by virus RNA extraction and RT-PCR. Then it was cloned in the expression vectors pGEX4T-1. The recombinant plasmid pGEX4T-1-VP8* was transformed to the E. coli BL21. Then the VP8*-GST fusion protein was expressed and purified by the affinity chromatograph. The protein of interest was validated by SDS-PAGE and Western Blot.
Results
The molecular weight of the VP8*-GST fusion protein was about 52 000 according to the SDS-PAGE. The bands of both 52 000 and 26 000 were shown in the Western Blot with the antibody against GST.
Conclusions
The LLR VP8* gene was obtained and cloned to the pGEX4T-1 vector. Moreover, the solvable VP8*-GST fusion protein was successfully expressed and purified.
Key words:
Rotavirus; LLR; VP8* Protein
科研通智能强力驱动
Strongly Powered by AbleSci AI