Preparation and Application of N Protein Polyclonal Antibody of Feline Infectious Peritonitis Virus SH2021 Strain
This study aimed to express the nucleocapsid protein(N)of feline infectious peritonitis virus(FIPV)strain SH2021 in vitro and prepare rabbit polyclonal antibodies.Primers were de-signed based on the N gene sequence of FIPV strain SH2021 to construct the recombinant expres-sion plasmid pCold-I-N.Subsequently,the recombinant plasmid was transformed into expres-sion-competent cells BL21(DE3),and expression was induced under conditions of 1.0 mmol·L-1 IPTG and 16 ℃.Finally,purification was performed using a His column,and the purified re-combinant protein was used to immunize New Zealand white rabbits to prepare polyclonal anti-bodies.The experimental results showed that the purified recombinant N protein had a size of ap-proximately 45 ku,and the titer of the prepared rabbit polyclonal antibodies against the N protein reached 1:512 000.The antibodies exhibited good reactivity and specificity towards the N pro-tein.This study successfully prepared rabbit polyclonal antibodies against the FIPV N protein,which showed good reactivity and specificity,providing important tools for FIPV antigen-anti-body detection and research on the biological functions of the N protein.
万方数据
维普
