佛山科学技术学院学报（自然科学版）2026，Vol.44Issue(3)：14-19,6.
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
Prokaryotic expression of porcine transmissible gastroenteritis virus N protein and preparation of its polyclonal antibody
李智丽 1王一臣 1伍俊杰 1文龙 1冉斌 1单同领2
作者信息
- 1. 佛山大学 动物科技学院,广东 佛山 528225
- 2. 中国农业科学院上海兽医研究所,上海 201100
- 折叠
摘要
Abstract
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.关键词
猪传染性胃肠炎病毒/N蛋白/原核表达:多克隆抗体Key words
TGEV/N protein/prokaryotic expression/polyclonal antibody分类
农业科技引用本文复制引用
李智丽,王一臣,伍俊杰,文龙,冉斌,单同领..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[J].佛山科学技术学院学报（自然科学版）,2026,44(3):14-19,6.
