猪传染性胃肠炎病毒N蛋白的原核表达及其鼠源多克隆抗体的制备OA

To prepare polyclonal antibody against the N protein of porcine transmissible gastroenteritis virus(TGEV),a fragment of the N gene was amplified by PCR using TGEV cDNA preserved in the laboratory as a template.This fragment was then cloned into the pCold TF expression vector after digestion and ligation,resulting in the construction of the recombinant plasmid pCold TF-TGEV-N.The plasmid was transformed into competent E.coli BL21,and the recombinant protein was purified by nickel column affinity chromatography following induced expression with IPTG.The purified protein was used as an immunogen to immunize BALB/c mice to produce polyclonal antibodies against the N protein.After the immunization program,the serum from the mice was collected and tested using I-ELISA.The results showed that the antibody titer exceeded 1∶100,000.The specificity of the antibodies was further confirmed by Western blot and IFA.The polyclonal antibodies specifically recognized the TGEV N protein and exhibited good reactivity and specificity to TGEV.