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来自溶组织内阿米巴强毒株的基因组和cDNA肌动蛋白序列。

Genomic and cDNA actin sequences from a virulent strain of Entamoeba histolytica.

作者信息

Edman U, Meza I, Agabian N

出版信息

Proc Natl Acad Sci U S A. 1987 May;84(9):3024-8. doi: 10.1073/pnas.84.9.3024.

Abstract

Invasiveness of Entamoeba histolytica strains that cause acute amoebiasis is characterized by aggressive behavior associated with cell motility and actin function. Analysis of actin genes from E. histolytica was initiated by devising methods for the isolation of biologically active nucleic acids, which allowed the preparation of cDNA and genomic DNA libraries. E. histolytica actin-encoding cDNAs and genomic clones have been isolated from libraries prepared from the virulent HM1:IMSS strain using a heterologous actin probe. Nucleotide sequence analysis of three independent cDNA clones and one genomic clone reveals a highly unusual codon bias and the absence of intervening sequences in E. histolytica actin. The coding sequence of the genomic clone is identical to that of two of the three cDNA clones. These represent at least two distinct mRNAs differing only by five silent changes in the protein coding sequence. Multiple genomic copies of the actin gene can be detected by Southern hybridization. E. histolytica actin exhibits a higher degree of homology to cytoplasmic than to muscle actin. Although the protein has been shown not to bind DNase I, the inferred amino acid sequence indicates conservation of all residues implied to participate in this binding.

摘要

导致急性阿米巴病的溶组织内阿米巴菌株的侵袭性特征在于与细胞运动性和肌动蛋白功能相关的侵袭性行为。通过设计用于分离生物活性核酸的方法,启动了对溶组织内阿米巴肌动蛋白基因的分析,这使得能够制备cDNA和基因组DNA文库。使用异源肌动蛋白探针,从有毒的HM1:IMSS菌株制备的文库中分离出了溶组织内阿米巴肌动蛋白编码cDNA和基因组克隆。对三个独立的cDNA克隆和一个基因组克隆的核苷酸序列分析揭示了溶组织内阿米巴肌动蛋白中高度不寻常的密码子偏好以及不存在间隔序列。基因组克隆的编码序列与三个cDNA克隆中的两个相同。这些代表了至少两种不同的mRNA,它们仅在蛋白质编码序列中有五个沉默变化。通过Southern杂交可以检测到肌动蛋白基因的多个基因组拷贝。与肌肉肌动蛋白相比,溶组织内阿米巴肌动蛋白与细胞质肌动蛋白具有更高程度的同源性。尽管已证明该蛋白质不结合DNase I,但推断的氨基酸序列表明参与这种结合的所有残基都具有保守性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/01b9/304793/62a324607af5/pnas00274-0470-a.jpg

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