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一种赋予酿酒酵母黏附与聚集能力的白色念珠菌DNA序列的分离

Isolation of a Candida albicans DNA sequence conferring adhesion and aggregation on Saccharomyces cerevisiae.

作者信息

Barki M, Koltin Y, Yanko M, Tamarkin A, Rosenberg M

机构信息

Maurice and Gabriela Goldschleger School of Dental Medicine, Tel-Aviv, Israel.

出版信息

J Bacteriol. 1993 Sep;175(17):5683-9. doi: 10.1128/jb.175.17.5683-5689.1993.

Abstract

Candida albicans is an opportunistic pathogen which may give rise to superficial and systemic infections. In the present study, C. albicans adhesion was studied by expression of C. albicans DNA sequences encoding adhesion functions in a nonadherent strain of Saccharomyces cerevisiae. Adherent transformant cells of S. cerevisiae harbouring a C. albicans genomic library cloned in a yeast-Escherichia coli shuttle vector were selected by using tissue culture-treated polystyrene as the attachment substratum. One transformant exhibited enhanced adhesion to treated and untreated polystyrene as well as autoaggregation, unlike control cells bearing the vector alone. Analysis of this clone revealed an insert of ca. 4.5 kb from C. albicans. Curing of the plasmid resulted in loss of adhesion and autoaggregation properties. A subclone bearing a reduced insert of 3.3 kb retained the ability to autoaggregate, to bind to treated and untreated polystyrene, and to adhere to buccal epithelial cells, unlike appropriate controls. Further subcloning of the insert to 2.7- and 1.9-kb fragments resulted in incremental decreases in adhesion and autoaggregation, whereas smaller fragments did not confer these properties. Hybridization of the 2.7-kb segment with C. albicans and S. cerevisiae DNA confirmed its origin as a single-copy sequence in the C. albicans genome as well as the absence of a homologous sequence in the genome of S. cerevisiae. The data suggest that the adhesion and aggregation phenomena of the transformant cells are related to expression of a C. albicans surface antigen encoded by the cloned DNA fragment.

摘要

白色念珠菌是一种机会致病菌,可引起浅表和全身感染。在本研究中,通过在非粘附性酿酒酵母菌株中表达编码粘附功能的白色念珠菌DNA序列来研究白色念珠菌的粘附。以经组织培养处理的聚苯乙烯作为附着基质,筛选出携带克隆于酵母-大肠杆菌穿梭载体中的白色念珠菌基因组文库的酿酒酵母粘附转化细胞。与仅携带载体的对照细胞不同,一个转化体对处理过和未处理过的聚苯乙烯表现出增强的粘附以及自身聚集能力。对该克隆的分析显示插入片段约为来自白色念珠菌的4.5 kb。质粒的消除导致粘附和自身聚集特性丧失。与适当的对照不同,一个携带3.3 kb减少插入片段的亚克隆保留了自身聚集、结合处理过和未处理过的聚苯乙烯以及粘附于颊上皮细胞的能力。将插入片段进一步亚克隆为2.7 kb和1.9 kb的片段导致粘附和自身聚集能力逐渐降低,而较小的片段则不具备这些特性。2.7 kb片段与白色念珠菌和酿酒酵母DNA的杂交证实其起源于白色念珠菌基因组中的单拷贝序列,并且在酿酒酵母基因组中不存在同源序列。数据表明,转化细胞的粘附和聚集现象与由克隆DNA片段编码的白色念珠菌表面抗原的表达有关。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8e87/206627/9d2f5c89cfea/jbacter00059-0400-a.jpg

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