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yin dao mao di chong ju shi xi bao qian yi yi zhi yin zi de yuan he biao da ji duo ke long kang ti zhi bei
Author(s): 
Pages: 17-20
Year: Issue:  1
Journal: Journal of Jiangsu University(Medicine Edition)

Keyword:  阴道毛滴虫巨噬细胞迁移抑制因子原核表达多克隆抗体;
Abstract: 目的:原核表达阴道毛滴虫巨噬细胞迁移抑制因子(macrophage migration inhibitory factor of Trichomonas vaginalis,Tv MIF)并制备多克隆抗体。方法:采用PCR技术扩增Tv MIF基因,并克隆至p TG19-T载体,酶切及测序鉴定后,再定向插入p ET-30a(+)载体。将重组载体转化至大肠埃希菌BL21(DE3)中,用IPTG诱导蛋白表达。采用镍柱亲和层析法纯化Tv MIF蛋白,并免疫新西兰白兔制备多克隆抗体,蛋白质印迹法检测多克隆抗体。结果:从阴道毛滴虫c DNA中扩增出Tv MIF基因片段,测序结果与基因库收录序列一致。成功构建p ET-30a-TvMIF重组质粒,重组Tv MIF蛋白约为15. 5 k Da,以可溶性蛋白形式存在。获得Tv MIF重组蛋白的多克隆抗体,蛋白印迹法检出Tv MIF的特异性条带。结论:成功获得阴道毛滴虫MIF蛋白并制备了多克隆抗体,可用于后续研究阴道毛滴虫致病机制。
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