摘要
为研究牛支原体(Mycoplasma bovis,Mb)硫氧还蛋白还原酶(thioredoxin reductase,TrxR)在EBL细胞内表达对其凋亡的影响,参照GenBank中Mb Hubei-1 TrxR序列设计引物,以Mb武威分离株基因组为模板,利用PCR扩增获得Mb TrxR基因,进而构建真核表达载体pVAX-TrxR和pEGFP-TrxR,并将其转染至EBL细胞内表达,然后用MTT法和流式细胞术分析Mb TrxR表达对EBL细胞增殖和凋亡的影响,用试剂盒检测分析EBL细胞中H_(2)O_(2)含量的变化。结果显示,Mb TrxR基因全长924 bp,成功在EBL细胞内表达;且该蛋白能促进EBL细胞增殖,抑制其凋亡;TrxR表达降低了EBL细胞内H_(2)O_(2)的含量。表明TrxR表达对EBL细胞的凋亡具有抑制作用,有利于EBL细胞的生长。研究结果为Mb TrxR生物学功能的深入研究积累了数据资料。
To study the effect of thioredoxin reductase(TrxR)expression of Mycoplasma bovis(Mb)on apoptosis in EBL cells,primers were designed according to the Mb Hubei-1 TrxR sequence in GenBank.The Mb TrxR gene was amplified by PCR from the genome of Mb Wuwei isolate.Eukaryotic expression vectors pVAX-TrxR and pEGFP-TrxR were constructed and transfected into EBL cells for expression.Then the effects of Mb TrxR expression on the proliferation and apoptosis of EBL cells were analyzed by MTT and flow cytometry,and the changes of H_(2)O_(2)content in EBL cells were analyzed by the kit.The results showed that the full-length 924 bp of Mb TrxR gene was successfully expressed in EBL cells,and the protein could promote the proliferation of EBL cells and inhibit its apoptosis.The expression of TrxR decreased the content of H_(2)O_(2)in EBL cells.The results showed that the expression of TrxR could inhibit the apoptosis of EBL cells and was beneficial to the growth of EBL cells.The results of this study accumulated data for the further study of the biological function of Mb TrxR.
作者
张立
何肖肖
马海云
赵云海
王青
蔡伟
邢小勇
武小椿
权国梅
温峰琴
包世俊
ZHANG Li;HE Xiao-xiao;MA Hai-yun;ZHAO Yun-hai;WANG Qing;CAI Wei;XING Xiao-yong;WU Xiao-chun;QUAN Guo-mei;WEN Feng-qin;BAO Shi-jun(College of Veterinary Medicine,Gansu Agricultural University,Lanzhou,Gansu 730070,China)
出处
《动物医学进展》
北大核心
2023年第9期29-37,共9页
Progress In Veterinary Medicine
基金
国家自然科学基金项目(32072863)。
