Prokaryotic expression and preparation of polyantibody of human histydyl-tRNA synthetase related gene.
- Author:
Xianfang MENG
1
;
Jing SHI
;
Xiaochun LIU
;
Jinzhong CHEN
Author Information
1. Department of Neurobiology, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China.
- Publication Type:Journal Article
- MeSH:
Antibodies;
genetics;
immunology;
Escherichia coli;
genetics;
metabolism;
Histidine-tRNA Ligase;
biosynthesis;
genetics;
immunology;
Humans;
Open Reading Frames;
genetics;
Prokaryotic Cells;
metabolism
- From:
Journal of Huazhong University of Science and Technology (Medical Sciences)
2004;24(6):535-555
- CountryChina
- Language:English
-
Abstract:
The aim of this study was to express and purify human histydyl-tRNA synthetase related gene and to prepare its polyantibody. The open reading frame was amplified by PCR, and then recombined into prokaryotic expression vector pQE30 and transformed into E. coli M15 for expression. The expressed products were induced by IPTG after the reconstructed pQE30 was transferred into M15. After purified by Ni affinity chromatography, the product was identified to be a single band by SDS-PAGE. The rabbits were inoculated with purified products. High-titer polyantibody was successfully prepared. Highly-purified expression product and prepared polyantibody may provide a good basis for further study.
