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变形链球菌中编码抗原A基因的分子克隆。

Molecular cloning of the Streptococcus mutans gene specifying antigen A.

作者信息

Dao M L, Chavez C, Hirachi Y, Ferretti J J

机构信息

Department of Pediatrics/St. Petersburg, University of South Florida, All Children's Hospital, St. Petersburg 33701.

出版信息

Infect Immun. 1989 Nov;57(11):3372-6. doi: 10.1128/iai.57.11.3372-3376.1989.

Abstract

A gene encoding a Streptococcus mutans surface protein antigen has been isolated from a strain GS-5 gene bank constructed via the Streptococcus-Escherichia coli shuttle vector pSA3. This E. coli recombinant clone, designated 4B2, expressed S. mutans proteins, as shown by Western immunoblot analysis with a specific rabbit antibody to S. mutans surface antigens. Three bands were observed, including a 52-kilodalton (kDa) protein (pI 5.7), a 29-kDa protein (pI 4.2), and a 20-kDa protein usually present in lower amounts. The 52- and 29-kDa proteins both reacted with a monoclonal antibody to S. mutans antigen A, a 29-kDa protein which has been characterized and used as a vaccine for the prevention of induced caries in rodents and monkeys. The 52-kDa protein, but not the 29-kDa protein, showed a capacity to bind to a broad number of carbohydrate polymers. The results from this study suggest that the recombinant 4B2 clone specifies a 52-kDa protein which is a precursor to the 29-kDa antigen A.

摘要

通过链球菌-大肠杆菌穿梭载体pSA3构建了变形链球菌GS-5菌株的基因文库,从中分离出了一个编码变形链球菌表面蛋白抗原的基因。用抗变形链球菌表面抗原的特异性兔抗体进行Western免疫印迹分析表明,这个命名为4B2的大肠杆菌重组克隆表达了变形链球菌蛋白。观察到三条带,包括一条52千道尔顿(kDa)的蛋白(等电点5.7)、一条29-kDa的蛋白(等电点4.2)以及一条含量通常较低的20-kDa蛋白。52-kDa和29-kDa的蛋白均与抗变形链球菌抗原A的单克隆抗体发生反应,抗原A是一种已被鉴定并用作预防啮齿动物和猴子龋齿疫苗的29-kDa蛋白。52-kDa的蛋白而非29-kDa的蛋白表现出与多种碳水化合物聚合物结合的能力。本研究结果表明,重组4B2克隆所表达的52-kDa蛋白是29-kDa抗原A的前体。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c34c/259826/b9bf32538964/iai00071-0126-a.jpg

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