期刊论文

Expression and analysis of the immune activity of the membrane protein MYPE2350of Mycoplasma penetrans

中文

中国病原生物学杂志

1673-5234

2021

16

10

1173-1177

10.13350/j.cjpb.211012

湖南教育厅优秀青年项目（No.19B496）； 湖南省自然科学基金项目（No.2020JJ4521）； 湖南省卫健委科学基金项目（No.20200706）； 衡阳市自然科学基金项目（No.2019-117）；

本校为第一机构

目的 对穿透支原体膜蛋白MYPE2350进行生物学信息学分析,构建重组原核表达质粒,表达、纯化并检测该蛋白的免疫学活性.方法 从NCBI网站获得穿透支原体特有MYPE2350蛋白的氨基酸序列,利用生物信息学软件预测和分析MYPE2350蛋白的理化性质、跨膜区、信号肽和B细胞表位等生物学特性.设计引物,PCR扩增MYPE2350基因片段,将其连接到质粒pET28a.将重组质粒pET28a-MYPE2350转化入大肠埃希菌BL21,用IPTG诱导重组MYPE2350蛋白的表达.分离包含体,使用尿素溶解后利用Ni2+亲和层析法纯化重组MYPE2350蛋白.用重组MYPE2350蛋白免疫小鼠,测定小鼠血清特异IgG含量和IgG2a/IgG1的比值,以及小鼠脾细胞分泌细胞因子的...

Objective To analyzed biological information on the membrane protein MYPE2350 of Mycoplasma penetrans, to construct a recombinant prokaryotic expression plasmid, to express and purify the recombinant MYPE2350 protein, and to test its immunological activity. Methods The amino acid sequence of the MYPE2350 protein of M. penetrans was obtained from the NCBI website. The physical and chemical properties, transmembrane domains, signal peptides, and B-cell epitopes of the MYPE2350 protein were analyzed using bioinformatic software. PCR primers were d...