Abstract: Anhui Province Key Laboratory of Veterinary Pathobiology and Disease Control predicted a new ETT2 chaperone molecule YgeG in the early stage. To obtain a large number of active YgeG protein, identify its function, and screen out its related active ETT2 secretion effector protein, the prokaryotic expression conditions of the target protein were optimized.The ygeG gene was PCR amplified, with the APEC 81 strain(AE81) as the template. The amplified product was cloned into pGEX-6p-1 prokaryotic expression vector, and the recombinant plasmid pGEX-6p-1-ygeG was constructed and identified by sequencing. Then, the correctly identified recombinant plasmid was transformed into E. coli BL21(DE3), and was induced by IPTG. And optimizing the induction conditions of the recombinant protein to determine the optimal conditions for the expression of YgeG recombinant protein. The results of SDS-PACE and Western Blot showed that the optimal induction conditions were determined to be an induction temperature of 16℃, an induction time of 16 h, and IPTG final concentration of 0.25 mmol/L. The size of the recombinant protein was about 44 ku, which was highly expressed in BL21(DE3) and mainly in the form of soluble protein and has good immunoreactivity and antigenicity. The AE81-YgeG protein was successfully expressed, which laid the foundation for the further study of the structure and function of the protein and ETT2, and provided a theoretical basis for the prevention and treatment of avian colibacillosis.

Key words: Escherichia coli, ygeG, ETT2, Prokaryotic expression, Protein purification

CLC Number: 

• S852.3

• Q78