阴道毛滴虫铁氧还蛋白基因的克隆与表达

发布时间：2018-01-09 02:25

本文关键词：阴道毛滴虫铁氧还蛋白基因的克隆与表达　出处：《四川大学》2005年硕士论文　论文类型：学位论文

【摘要】：研究背景及目的:阴道毛滴虫(Trichomonas vaginalis)是寄生在人体泌尿生殖道的一种常见病原体,可引起阴道毛滴虫病(Xrichomoniasis)。自1836年Donné首次描述该虫的特点及所致疾病的症状以来,至今仍是威胁人类健康的主要的性传播疾病之一。铁氧还蛋白(Ferredoxin Fd)是自然界分布很广的低分子量铁硫蛋白,最常见的是[2Fe-2S]Fd,在阴道毛滴虫中它是氢化酶体(Hydrogenosome)电子传递链的主要成分,它参与丙酮酸盐:铁氧还蛋白氧化还原酶(pyruvate:ferredoxin oxidoreductase PFOR)介导的丙酮酸盐氧化脱羧形成乙酰辅酶A和ATP反应,是此过程中的电子供体,通过电子传递参与了—线抗滴虫药物甲硝唑(metronidazole)在体内的活化。尽管应用甲硝唑已经数十年,其抗药性的发展还是较缓慢。然而,近年来临床上甲硝唑抗性株的发现和不断增加,使得对甲硝唑在体内的作用机制和抗药性的发展的研究就尤为重要。Quon和Johnson等发现滴虫抗性株细胞中Fd明显低于敏感株,Northern杂交分析,Fd的mRNA水平下降50-65%,基因转录下降40-65%,Land等通过实验发现药物高度抵抗的虫株内Fd mRNA的水平下降90%,免疫印记显示,在高度抵抗株内几乎检测不到PFOR和Fd,甲硝唑在抗药株内活化减少。本课题对Fd基因及蛋白质的研究将有助于阐明阴道毛滴虫抗药株抗性机制。本研究分两个部分,第一部分:目的:Fd基因的克隆。方法:从我国四川地区临床分离阴道毛滴虫(阴道毛滴虫四川分离株—1),应用螯合树脂
[Abstract]:Background and objective: Trichomonas vaginalis is a common pathogen in human genitourinary tract. Trichomonas vaginalis can cause trichomonas vaginalis. Since 1836, Donn 茅 first described the characteristics and symptoms of the disease. Ferredoxin Fd is one of the major sexually transmitted diseases that threaten human health. Ferredoxin Fd is a widely distributed low molecular weight ferrithionein in nature, the most common of which are. [2Fe-2S] FD, which is the main component of the electron transport chain of Hydrogenosome in Trichomonas vaginalis. It participates in pyruvate: ferredoxin oxidoreductase redoxin pyruvate: ferredoxin, pyruvate: ferredoxin, pyruvate: ferredoxin). The oxidative decarboxylation of pyruvate into acetyl coenzyme A and ATP reaction was mediated. Metronidazole is the electron donor involved in the activation of metronidazole, a novel antitrichomonium drug, although metronidazole has been used for decades. The development of resistance to metronidazole is still slow. However, in recent years, metronidazole resistant strains have been found and increased. It is very important to study the mechanism of metronidazole in vivo and the development of drug resistance. Quon and Johnson showed that the FD of trichomonas resistant strain was significantly lower than that of susceptible strain. Northern blot analysis showed that the mRNA level of Fd decreased by 50-65%, and the transcription of Fd gene decreased by 40-65%. Land et al. found that the level of FD mRNA in the highly resistant strains decreased 90%. The immunological imprinting showed that PFOR and FD could hardly be detected in the highly resistant strains. Metronidazole activates less in resistant strains. The study of FD gene and protein will be helpful to elucidate the resistance mechanism of Trichomonas vaginalis resistance strain. This study is divided into two parts. Part one: to clone the gene of: Fd. Methods: to isolate Trichomonas vaginalis (Trichomonas vaginalis) (Trichomonas vaginalis strain-1) from the clinic in Sichuan, china, and apply chelating resin
【学位授予单位】：四川大学
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：R383

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