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白色念珠菌假定膜蛋白编码新基因CSP37的克隆与特性分析

Cloning and characterization of CSP37, a novel gene encoding a putative membrane protein of Candida albicans.

作者信息

Sentandreu M, Nieto A, Iborra A, Elorza M V, Ponton J, Fonzi W A, Sentandreu R

机构信息

Seccio Departamental de Microbiologia, Facultat de Farmàcia, Universitat de València, Spain.

出版信息

J Bacteriol. 1997 Aug;179(15):4654-63. doi: 10.1128/jb.179.15.4654-4663.1997.

Abstract

In the course of an analysis of the functions and assembly of the cell wall of Candida albicans, we have cloned and characterized a gene, which we designated CSP37 (cell surface protein), encoding a 37-kDa polypeptide which is a membrane-associated protein. The gene was isolated by immunological screening of a DNA library constructed from mycelial cells with a polyclonal serum raised against cell walls of this morphology. Analysis of the nucleotide sequence of a corresponding genomic DNA fragment revealed a single open reading frame which encodes a predicted protein of 321 amino acids with no significant homology to others in the databases. Disruption of the CSP37 gene by the method described by Fonzi and Irwin (Genetics 134:717-728, 1993) eliminated expression of the Csp37 protein. The mutant strains showed no apparent defect in cell viability, growth, or cell wall assembly but displayed attenuated virulence in systemic infections induced in mice and reduced the ability to adhere to polystyrene.

摘要

在对白色念珠菌细胞壁的功能和组装进行分析的过程中,我们克隆并鉴定了一个基因,将其命名为CSP37(细胞表面蛋白),该基因编码一种37 kDa的多肽,是一种膜相关蛋白。通过用针对这种形态细胞壁产生的多克隆血清对由菌丝体细胞构建的DNA文库进行免疫筛选,分离出了该基因。对相应基因组DNA片段的核苷酸序列分析显示有一个单一的开放阅读框,其编码一个预测的由321个氨基酸组成的蛋白质,该蛋白质与数据库中的其他蛋白质没有明显的同源性。采用Fonzi和Irwin(遗传学134:717 - 728, 1993)描述的方法破坏CSP37基因后,消除了Csp37蛋白的表达。突变菌株在细胞活力、生长或细胞壁组装方面没有明显缺陷,但在小鼠体内诱导的全身感染中表现出毒力减弱,并且降低了黏附于聚苯乙烯的能力。

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