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克氏锥虫基因组包含与利什曼原虫质子泵极为相似的离子动力ATP酶基因。

The Trypanosoma cruzi genome contains ion motive ATPase genes which closely resemble Leishmania proton pumps.

作者信息

Meade J C, Li C, Stiles J K, Moate M E, Penny J I, Krishna S, Finley R W

机构信息

Department of Microbiology, University of Mississippi Medical Center, 2500 North State Street, Jackson, MS 39216-4505, USA.

出版信息

Parasitol Int. 2000 Dec;49(4):309-20. doi: 10.1016/s1383-5769(00)00061-1.

Abstract

DNA fragments homologous to members of the family of P-type ion-motive ATPases were identified in Trypanosoma cruzi by polymerase chain reaction (PCR) amplification. The sequence of one fragment, which closely resembled (87% identity) the tandemly linked proton pumps in Leishmania, was used to characterize the H(+)-ATPase genes in T. cruzi. The T. cruzi proton pump locus contains four tandemly repeated genes (TCH1-4) separated by 1.1 kb intergenic regions. The nucleotide sequence of one cloned gene of the tandem array contains a 2775 nt open reading frame encoding a predicted 101908-Da protein of 925 amino acids. The TCH genes are expressed as 3.8 and 4.9 kb polyadenylated transcripts in the epimastigote stage; expression of both transcripts is reduced in metacyclic trypomastigotes. Results of 5' and 3' RACE transcript mapping indicate that the 3.8 kb message is generated from within the tandemly repeated locus. The 3.8 kb TCH transcript has the T. cruzi mini-exon appended to a short (40 nt) 5' untranslated region (UTR) and has a 927 nt 3' UTR. The full peptide sequence of the T. cruzi proton pump is 80% identical to the Leishmania pump but lacks the extended carboxyl tail present in the Leishmania ATPase. An antibody that recognizes the 110-kDa Leishmania donovani proton pump cross-reacts with a 100-kDa protein in lysates of T. cruzi epimastigotes.

摘要

通过聚合酶链反应(PCR)扩增,在克氏锥虫中鉴定出了与P型离子驱动ATP酶家族成员同源的DNA片段。其中一个片段的序列与利什曼原虫中串联连接的质子泵高度相似(同一性为87%),被用于表征克氏锥虫中的H(+)-ATP酶基因。克氏锥虫质子泵基因座包含四个串联重复基因(TCH1-4),间隔1.1 kb的基因间区域。串联阵列中一个克隆基因的核苷酸序列包含一个2775 nt的开放阅读框,编码一个预测的925个氨基酸、101908 Da的蛋白质。TCH基因在无鞭毛体阶段表达为3.8 kb和4.9 kb的多聚腺苷酸化转录本;在循环后期锥鞭毛体中,这两种转录本的表达均降低。5'和3' RACE转录本图谱分析结果表明,3.8 kb的信息来自串联重复基因座内部。3.8 kb的TCH转录本在短的(40 nt)5'非翻译区(UTR)后附加了克氏锥虫小外显子,并有一个927 nt的3' UTR。克氏锥虫质子泵的完整肽序列与利什曼原虫泵的同一性为80%,但缺少利什曼原虫ATP酶中存在的延长羧基末端。一种识别110-kDa杜氏利什曼原虫质子泵的抗体与克氏锥虫无鞭毛体裂解物中的一种100-kDa蛋白质发生交叉反应。

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