Chaperone protein facilitates soluble expression of Mannheimia haemolytica PlpE protein in Escherichia coli
10.16303/j.cnki.1005-4545.2025.07.05
- VernacularTitle:伴侣蛋白促使溶血性曼氏杆菌PlpE蛋白在大肠杆菌中的可溶性表达
- Author:
Yuemei ZHANG
1
;
Na WANG
;
Lingli DAI
;
Fan ZHANG
;
Yue SONG
;
Shihua ZHAO
;
Jingyu SHI
;
Wenhua GUO
;
Shengjie SU
;
Fan BAI
Author Information
1. 内蒙古自治区农牧业科学院,内蒙古呼和浩特 010031
- Publication Type:Journal Article
- Keywords:
molecular chaperones;
Mannheimia haemolytica;
Pasteurella lipoprotein E;
soluble ex-pression;
prokaryotic expression system
- From:
Chinese Journal of Veterinary Science
2025;45(7):1388-1393
- CountryChina
- Language:Chinese
-
Abstract:
To verify whether chaperones can promote the soluble expression of PlpE in Escherichia coli and whether the expressed protein is active,prokaryotic expression and Western blot detection were performed.The results showed that:The PlpE prokaryotic expression vector pET-32a(+)-plpE was expressed as inclusion body,and the expression form was not changed by changing the concentration of inducer,induction time and temperature.The companion proteins pG-KJE8,pGro7,pKJE7 and pG-Tf2 were co-expressed with pET-32a(+)-plpE in Eschierichia coli expres-sion system,respectively.When the final concentration of IPTG of 0.5 mmol/L,L-arabinose of 0.5 g/L or tetracycline of 5.0 μg/L were added as inducers and induced at 37 ℃ for 8 h,the results showed that the molecular companion pGro7 could change the expression of rp-PlpE from inclu-sion body to soluble expression.pG-KJE8,pKJE7 and pG-Tf2 had no effect on the expression of rp-PlpE.The soluble rp-PlpE can react specifically with the positive serum of Mannheimia haemolyti-ca.Therefore,the study showed that the co-expression of the chaperone protein pGro7 can make the rp-PlpE protein express in a soluble form,and the purified protein exhibits reactogenicity.These findings lay the foundation for the establishment of a subunit vaccine and serological diagno-sis methods for Mannheimia haemolytica.
