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溶组织内阿米巴铁氧化还原蛋白基因的核苷酸序列分析

Nucleotide sequence analysis of an Entamoeba histolytica ferredoxin gene.

作者信息

Huber M, Garfinkel L, Gitler C, Mirelman D, Revel M, Rozenblatt S

机构信息

Department of Biophysics, Weizmann Institute of Science, Rehovot, Israel.

出版信息

Mol Biochem Parasitol. 1988 Oct;31(1):27-33. doi: 10.1016/0166-6851(88)90142-9.

Abstract

A cDNA clone (subclone B) previously isolated from the human parasite Entamoeba histolytica was characterized. DNA sequence analysis of subclone B identified the DNA as that encoding apoferredoxin. E. histolytica ferredoxin cDNA contains unusually short 5' and 3' noncoding regions of 9 and 25 nucleotides, respectively. A genomic ferredoxin clone was isolated from E. histolytica DNA, and comparison of genomic and cDNA sequences revealed that the ferredoxin gene is unspliced. The deduced amino acid sequence of E. histolytica ferredoxin resembles clostridial type of ferredoxins, and shows an arrangement of cysteines characteristic for the coordination of 2[4Fe-4S] centres. Of interest is the absence of an aromatic amino acid in the N-terminal region of the protein, a feature which is conserved in clostridial ferredoxins. Southern blot analysis of three different E. histolytica strains (200:NIH, Rahman and HM-1:IMSS) demonstrated the presence of a family of at least two ferredoxin genes. One of these genes is marked by restriction length polymorphisms in different strains of E. histolytica.

摘要

对先前从人体寄生虫溶组织内阿米巴中分离出的一个cDNA克隆(亚克隆B)进行了特性分析。亚克隆B的DNA序列分析确定该DNA编码脱辅基铁氧化还原蛋白。溶组织内阿米巴铁氧化还原蛋白cDNA分别含有异常短的5'和3'非编码区,长度分别为9个和25个核苷酸。从溶组织内阿米巴DNA中分离出一个基因组铁氧化还原蛋白克隆,基因组序列与cDNA序列的比较显示铁氧化还原蛋白基因未发生剪接。溶组织内阿米巴铁氧化还原蛋白推导的氨基酸序列类似于梭菌型铁氧化还原蛋白,并显示出用于2个[4Fe-4S]中心配位的半胱氨酸排列特征。有意思的是,该蛋白质N端区域没有芳香族氨基酸,这一特征在梭菌铁氧化还原蛋白中是保守的。对三种不同的溶组织内阿米巴菌株(200:NIH、拉赫曼和HM-1:IMSS)进行的Southern印迹分析表明存在一个至少由两个铁氧化还原蛋白基因组成的家族。这些基因中的一个在不同的溶组织内阿米巴菌株中表现出限制性片段长度多态性。

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