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tvcp12: a novel Trichomonas vaginalis cathepsin L-like cysteine proteinase-encoding gene

机译:TVCP12:一种新型滴虫滴虫阴道阴茎L样半胱氨酸蛋白酶编码基因

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Trichomonas vaginalis is the causative agent of trichomoniasis, one of the most common sexually transmitted diseases in humans. This protozoan has multiple proteinases that are mainly of the cysteine proteinase (CP) type, some of which are known to be involved in the parasite's virulence. Here, a novel T. vaginalis CP-encoding gene, tvcp12, was identified and characterized. tvcp12 is 948?bp long and encodes a predicted 34·4?kDa protein that has the characteristics of the papain-like CP family. TvCP12 does not appear to have a signal peptide, suggesting that this is a cytoplasmic CP. By Southern blot assays, the tvcp12 gene was found as a single copy in the T. vaginalis genome. Remarkably, Northern blot experiments showed a single transcript band of ~1·3?kb in the mRNA obtained from parasites grown in low iron conditions and no transcript was observed in the mRNA from parasites grown in high iron conditions. By RT-PCR assays, a 270?bp band was amplified from the cDNA of parasites grown in low iron medium, which was very faint when cDNA from parasites grown in high iron conditions was used. Transcripts of the 3′ region obtained in both iron conditions presented differences in their poly(A) tail length. These data suggest that tvcp12 is another gene that is negatively regulated by iron and that the length of the poly(A) tail may be one of the factors involved in the iron-modulated protein expression.
机译:流动性滴虫是滴虫病的致病因子,人类中最常见的性传播疾病之一。该原生动物具有多种蛋白酶,主要是半胱氨酸蛋白酶(CP)型,其中一些已知参与寄生虫的毒力。这里,鉴定并表征了一种新的T.阴道CP编码基因TVCP12。 TVCP12是948?BP长,编码预测的34·4?KDA蛋白,其具有木瓜蛋白酶样CP系列的特征。 TVCP12似乎没有信号肽,表明这是一种细胞质CP。通过Southern印迹测定,在阴道基因组中发现了TVCP12基因作为单一拷贝。值得注意的是,Northern印迹实验显示,在低铁条件下生长的寄生虫中获得的mRNA中的单个转录条带〜1·3?Kb,并且在高铁条件下生长的寄生虫中未观察到转录物。通过RT-PCR测定,从低铁介质中生长的寄生虫的cDNA扩增270〜BP带,当使用高铁条件生长的寄生虫CDNA时非常微弱。在两种铁条件下获得的3'区域的转录物呈现它们的聚(a)尾长的差异。这些数据表明,TVCP12是由熨斗产生负面调节的另一个基因,并且聚(a)尾部的长度可以是铁调制蛋白表达中涉及的因素之一。

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