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黏性放线菌抗原在大肠杆菌中的表达:2型菌毛结构基因(fimA)的克隆

Expression of Actinomyces viscosus antigens in Escherichia coli: cloning of a structural gene (fimA) for type 2 fimbriae.

作者信息

Donkersloot J A, Cisar J O, Wax M E, Harr R J, Chassy B M

出版信息

J Bacteriol. 1985 Jun;162(3):1075-8. doi: 10.1128/jb.162.3.1075-1078.1985.

Abstract

A cosmid gene library of Actinomyces viscosus T14V was prepared in Escherichia coli to examine the expression of A. viscosus antigens and to gain insight into the structure of A. viscosus type 1 and type 2 fimbriae. Out of this library of 550 clones, 28 reacted in a colony immunoassay with antibodies against A. viscosus cells. The proteins responsible for these reactions were identified in three clones. Clones AV1209 and AV2009 displayed nonfimbrial antigens with subunits of 40 and 58 kilodaltons, respectively. Clone AV1402 showed a 59-kilodalton protein that reacted with monospecific antibody against type 2 fimbriae and that comigrated with a subunit of type 2 fimbriae during sodium dodecyl sulfate-polyacrylamide gel electrophoresis. This indicates that AV1402 expresses a gene (fimA) for a subunit of A. viscosus type 2 fimbriae.

摘要

构建了粘性放线菌T14V的黏粒基因文库,该文库存在于大肠杆菌中,用于检测粘性放线菌抗原的表达,并深入了解粘性放线菌1型和2型菌毛的结构。在这个包含550个克隆的文库中,有28个在菌落免疫测定中与抗粘性放线菌细胞的抗体发生反应。在三个克隆中鉴定出了引发这些反应的蛋白质。克隆AV1209和AV2009分别显示出具有40和58千道尔顿亚基的非菌毛抗原。克隆AV1402显示出一种59千道尔顿的蛋白质,它与抗2型菌毛的单特异性抗体发生反应,并且在十二烷基硫酸钠-聚丙烯酰胺凝胶电泳期间与2型菌毛的一个亚基迁移率相同。这表明AV1402表达了一个编码粘性放线菌2型菌毛亚基的基因(fimA)。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8983/215885/0ff891533fa7/jbacter00223-0228-a.jpg

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