A prokaryotic expression vector TLN-58 and its application method for antibacterial activity against pathogenic bacteria in the digestive tract of raccoon dogs

Abstract

The present invention discloses a prokaryotic expression vector TLN-58 and its application for antibacterial activity against pathogenic bacteria in the digestive tract of raccoon dogs. In this invention, purified protein was obtained through a prokaryotic expression system, and polyclonal antibodies were produced. The in vitro antibacterial efficacy and stability of the purified protein against Escherichia coli and Salmonella derived from raccoons were systematically determined. Using genetic recombination technology, the TLN-58 gene fragment was cloned into the vector pGEX-4T-1, successfully generating the recombinant plasmid pGEX-4T-1-TLN-58. After transformation of this vector into E. coli codon plus strains, IPTG induction enabled efficient expression of the target protein in soluble form.This approach bypassed the complex steps associated with refolding inclusion bodies, thereby simplifying subsequent purification procedures. Simultaneously, polyclonal antibodies produced by immunizing New Zealand White rabbits with the purified protein exhibited high titers and excellent specificity. These antibodies meet the requirements for protein identification and activity validation, providing a reliable tool for relevant research.