中国动物传染病学报2025，Vol.33Issue(3)：42-50,9.
鸡毒支原体磷酸甘油酸变位酶的亚细胞定位及免疫原性分析
Subcellular Localization and Immunogenicity of Phosphoglycerate Mutase(PGM)in Mycoplasma gallisepticum
摘要
Abstract
Mycoplasma gallisepticum(MG)infects avian such as chickens and ducks,causing chronic respiratory diseases,decrease of laying rate,growth and development retardation and resulting in great economic losses to the poultry industry.Phosphoglycerate mutase(PGM)is an important enzyme involved in glycolysis and gluconeogenesis.Some studies have shown that PGM is distributed on the membrane surface of some pathogenic bacteria and binds to extracellular proteins of host cells,playing an important role in the process of pathogenesis.However,there are few researches about MGPGM.A series of analysis including prokaryotic expression,subcellular localization and the immunogenicity of MGPGM were carried out in this study to pave the way for the further exploration of the biological functions of MGPGM.The full length of MGPGM gene was amplified by PCR and inserted into the expression vector pET-28a(+).Then the recombinant MGPGM(rMGPGM)protein was expressed in Escherichia coli BL21(DE3)and purified.The whole cells,membrane and cytoplasmic proteins were extracted and the subcellular localization of PGM in MG was analyzed by Western blotting with the pre-prepared rabbit anti-rMGPGM sera.Then the localization of PGM on the membrane surface of MG was determined by immunofluorescence analysis.Furthermore,MG-infected chicken serum was used to react with rMGPGM protein in Western blot to determine its immunogenicity.The recombinant rMGPGM protein was expressed in Escherichia coli BL21 and the titer of the rabbit anti-rMGPGM was 102 400.The rabbit anti-rMGPGM serum reacted with MG whole cells,membrane and cytoplasmic proteins in Western blot.The PGM was abundant in cytoplasm and partially present on the membrane.Suspension immunofluorescence assay further determined that the PGM was on the surface of MG.In addition,the MGPGM had a significant reaction band with MG-infected chicken serum,indicating it had good immunogenicity.This study confirmed that PGM was distributed on the membrane surface of MG and had good immunogenicity.These results provided reference value for the further study of PGM in the pathogenesis of MG.关键词
鸡毒支原体/磷酸甘油酸变位酶/原核表达/亚细胞定位/免疫原性Key words
Mycoplasma gallisepticum/phosphoglycerate mutase/prokaryotic expression/subcellular localization/immunogenicity分类
农业科技引用本文复制引用
赵宇馨,于圣青,祁晶晶,尚原冰,李浩然,刘婷,王宇,王少辉,田明星,高崧..鸡毒支原体磷酸甘油酸变位酶的亚细胞定位及免疫原性分析[J].中国动物传染病学报,2025,33(3):42-50,9.基金项目
国家自然科学基金(31902244) （31902244）
中央级公益性科研院所基本科研业务费专项资金(2022JB05) （2022JB05）
国家重点研发项目(2017YFD0500705) （2017YFD0500705）
