摘要
目的-将阴道毛滴虫铁氧还蛋白(ferredoxin,Fd)基因在大肠埃希菌中诱导表达。方法-制备阴道毛滴虫可溶性抗原,多点注射免疫家兔,获得的血清用ELISA测定其抗体效价。将原核表达重组质粒pET3C-Fd转化入大肠埃希菌BL21(DE3)感受态细胞中,异丙基-β-D-硫代半乳糖苷(IPTG)诱导蛋白质表达。结果-制备出抗阴道毛滴虫多克隆抗体,抗体效价在1∶8000以上,用于免疫印迹实验。经十二烷基磺酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)和免疫印迹(Western blot)分析,重组质粒在大肠埃希菌中表达出Fd。结论-在大肠埃希菌中表达出了Fd。
Objective Ferredoxin gene of Trichomonas vaginalis was induced to express in E. coli. Methods Antiserum against T. vaginslis was generated in rabbits injected subcutaneously and intramuscularly with ,soluble antigen. Antibody titre was determined with ELISA method. The recombinant prokaryotic plasmid pET3C-Fd was transferred into E. coli BL21 （DE3）. The ferredoxin was induced to express by IPTG. Results The polyclonal antibody obtained from rabbit had a titre of above 1 ： 8000 and the antibody can be used as primary antibody of Western blot assay. The expressed protein in E. coli was identified by SDS-PAGE and Western blot. Conclusion The ferredoxin was successfully expressed in E. coli.
出处
《四川动物》
CSCD
北大核心
2006年第4期692-694,共3页
Sichuan Journal of Zoology
关键词
阴道毛滴虫
铁氧还蛋白
多克隆抗体
基因表达
Trichomonas vaginalis
ferredoxin
polyclonal antibidy
gene expression
