溶组织内阿米巴：肌动蛋白cDNA的克隆与特性分析

Entamoeba histolytica: cloning and characterization of actin cDNA.

作者信息

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

出版信息

Mol Biochem Parasitol. 1987 Jul;24(3):227-35. doi: 10.1016/0166-6851(87)90154-x.

Abstract

In order to study gene expression in the human parasite Entamoeba histolytica, a cDNA library of E. histolytica strain 200:NIH was constructed using the phage vector lambda gt10. Three cDNA clones (A, B and C) were selected for further analysis. Each of the three clones hybridized to a distinct mRNA. Two of these mRNAs were translated in vitro after hybrid selection, and yielded distinct translation products. One of these mRNAs, selected by hybridization to clone A, encodes the most abundantly expressed protein in E. histolytica. DNA sequence analysis of this cDNA clone identified the DNA as that encoding actin. The deduced amino acid sequence of E. histolytica actin resembles both cytoplasmic and muscle actins and has an unusual N-terminal glycine residue. We have shown that a family of actin genes is present in E. histolytica. Six different E. histolytica actin clones were obtained from a lambda gt10 genomic library using subcloned cDNA probes. Southern analysis of three different E. histolytica strains (200:NIH, Rhaman, and HM-1:IMSS) revealed at least four different actin genes. Strain HM-1:IMSS, however, differs by the presence of an additional actin gene.

摘要

为了研究人类寄生虫溶组织内阿米巴的基因表达，使用噬菌体载体λgt10构建了溶组织内阿米巴200:NIH菌株的cDNA文库。选择了三个cDNA克隆（A、B和C）进行进一步分析。这三个克隆中的每一个都与一种独特的mRNA杂交。其中两个mRNA在杂交筛选后进行了体外翻译，并产生了独特的翻译产物。通过与克隆A杂交选择的其中一种mRNA编码溶组织内阿米巴中表达最丰富的蛋白质。对该cDNA克隆的DNA序列分析确定该DNA编码肌动蛋白。溶组织内阿米巴肌动蛋白推导的氨基酸序列既类似于细胞质肌动蛋白，也类似于肌肉肌动蛋白，并且有一个不寻常的N端甘氨酸残基。我们已经表明溶组织内阿米巴中存在一个肌动蛋白基因家族。使用亚克隆的cDNA探针从λgt10基因组文库中获得了六个不同的溶组织内阿米巴肌动蛋白克隆。对三种不同的溶组织内阿米巴菌株（200:NIH、Rhaman和HM-1:IMSS）进行的Southern分析揭示了至少四个不同的肌动蛋白基因。然而，HM-1:IMSS菌株因存在一个额外的肌动蛋白基因而有所不同。