中国动物传染病学报2023，Vol.31Issue(5)：55-63,9.
一株滑液囊支原体的分离鉴定及其磷酸丙糖异构酶(Tpi)和丙酮酸激酶(PK)抗体制备
Isolation and Identification of a Mycoplasma Synovialis Strain and Preparation of Its Triose Phosphate Isomerase(Tpi)and Lian Pyruvate Kinase(Pk)Antibodies
摘要
Abstract
Triosephosphate isomerase(Tpi)and pyruvate kinase(PK)are the key enzymes for glycolysis of Mycoplasma synoviae(MS).Tpi and PK of M.bovine and M.gallisepticum are located on the cell membranes and cytoplasms,respectively,which participate in adhesion to host cells.However,tpiA and pyk of M.synoviae have not been reported yet.The growth characteristics of M.synoviumisolated from a chicken farm in Shandong province were investigated in the present study.Then the point mutations of TpiA and Pyk were amplified by overlap PCR method,which were respectively linked to pET-28a expression vector and expressed in BL21(DE3).The recombinant proteins of TpiA and Pyk were used to immunize New Zealand rabbits,respectively.The results showed that the highest growth titration of the M.synoviae isolates MS-SD2020 was 109 at 60 h of growth.Sequence analysis of TpiA and Pyk showed that both were highly conserved in different M.synoviae species.The recombinant proteins of TpiA and Pyk with the sizes of 35.28 kDa and 63.36 kDa were successfully expressed,and the rabbit serum antibody titers were 32,000 and 2,048,000,respectively.This study provided a reference for the further study on the function of TpiA and Pyk in M.synoviae.关键词
滑液囊支原体/磷酸丙糖异构酶/丙酮酸激酶/抗体制备Key words
Mycoplasma synoviae/triosephosphate isomerase/pyruvate kinase/antibody preparation分类
农业科技引用本文复制引用
连丽燕,巩新民,郭龙宗,李玉燕,蒋蔚,陈兆国,祁克宗,魏晓峰,韩先干..一株滑液囊支原体的分离鉴定及其磷酸丙糖异构酶(Tpi)和丙酮酸激酶(PK)抗体制备[J].中国动物传染病学报,2023,31(5):55-63,9.基金项目
上海科委实验动物专项(18140900700) （18140900700）
