摘要
为研究鸡毒支原体(MG)的可变脂蛋白血凝素(VLHA)的生物学功能,对VLHA 3.03蛋白进行原核表达,然后进行了免疫原性和免疫反应性分析。通过Overlap PCR方法对MG Rlow株的vlha 3.03基因进行点突变扩增,将突变成功的vlha 3.03基因全长与原核表达载体pCold Ⅰ连接,连接产物转入大肠杆菌BL21进行诱导表达和纯化。用纯化后的重组蛋白rVLHA3.03免疫新西兰大白兔,制备兔多克隆抗体,并用ELISA检测抗体效价,再用制备的兔多克隆抗体、MG不同分离株阳性鸡血清及MS标准阳性血清分别与rVLHA3.03蛋白进行Western blot反应,分析rVLHA3.03蛋白的免疫原性和免疫反应性。结果表明,在大肠杆菌BL21中成功表达rVLHA3.03蛋白,且该蛋白具有较好的免疫原性和免疫反应性。
In order to investigate the biological function of the variable lipoprotein hemagglutinin(VLHA)of Mycoplasma gallisepticum(MG),the recombinant VLHA3.03 protein was prokaryotically expressed for its immunogenicity and immunoreactivity analysis.The vlha 3.03 gene of MG Rlow strain was amplified by overlap PCR to obtain point mutations,then the mutant vlha 3.03 gene was ligated to the expression vector pCold Ⅰ and then transformed into E.coli BL21.The rabbit antiserum against rVLHA3.03 was prepared by immunizing New Zealand white rabbits with purified rVLHA3.03 protein and the antibody titer was detected in ELISA.The rVLHA3.03 protein was recognized in Western blot by rabbit anti-rVLHA3.03 serum,the positive chicken sera of different MG isolates and MSpositive serum,suggesting its good immunogenicity and immunoreactivity.
作者
陈玥彤
祁晶晶
胡增金
赵宇馨
李浩然
刘晓涵
王少辉
田明星
郭伶
于圣青
周铁忠
CHEN Yuetong;QI Jingjing;HU Zengjin;ZHAO Yuxin;LI Haoran;LIU Xiaohan;WANG Shaohui;TIAN Mingxing;GUO Ling;YU Shengqing;ZHOU Tiezhong(College of Animal Husbandry and Veterinary Medicine,Jinzhou Medicine University,Jinzhou 121000,China;Shanghai Veterinary Research Institute,CAAS,Shanghai 200241,China;College of Animal Science and Technology,Anhui Agricultural University,Hefei 230036,China)
出处
《中国动物传染病学报》
CAS
北大核心
2022年第2期134-141,共8页
Chinese Journal of Animal Infectious Diseases
基金
国家自然科学基金(31902244)
国家重点研发项目(2017YFD0500705)
辽宁省科学技术计划项目(2021JH2/10200009)。
关键词
鸡毒支原体
VLHA
原核表达
免疫原性
免疫反应性
Mycoplasma galliscepticum
VLHA
prokaryotic expression
immunogenicity
immunoreactivity
