Cloning and expression of an adhesin antigen of Streptococcus sanguis G9B in Escherichia coli.
Cloning and expression of an adhesin antigen of Streptococcus sanguis G9B in Escherichia coli.
复制标题
血链球菌 G9B 粘附素抗原在大肠杆菌中的克隆和表达。
DOI：
10.1099/00221287-135-3-531
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发表时间：
1989
期刊：
影响因子：
--
通讯作者：
Demuth,DR
中科院分区：
文献类型：
--
作者：
Rosan,B;Baker,CT;Nelson,GM;Berman,R;Lamont,RJ;Demuth,DR
构建了血链球菌 G9B 菌株的基因组文库，并使用 λgt11 表达载体在大肠杆菌中进行表达。用多克隆抗 G9B IgG 探测扩增的文库，分离出 13 个抗原阳性克隆。一个克隆的裂解物（命名为 PP39）吸收了抗 G9B IgG 的粘附抑制活性。该克隆包含大约 2000 bp 的插入片段，并表达与单特异性抗粘附素抗体反应的独特的 200 和 53 kDa 蛋白质。 200 kDa 的蛋白也与抗β-半乳糖苷酶 IgG 发生反应，表明它是融合蛋白，其中 84 kDa 代表链球菌粘附素。 84 和 53 kDa 蛋白的大小与链球菌抗原复合物中的主要多肽相似，该复合物与 G9B 与唾液涂覆的羟基磷灰石的粘附有关。 53 kDa 片段可能由重组多肽的翻译后切割产生。
A genomic library ofStreptococcus sanguis, strain G9B, was constructed and expressed inEscherichia coliusing a λgt11 expression vector. The amplified library was probed with polyclonal anti-G9B IgG and 13 antigen-positive clones were isolated. A lysate of one clone, designated PP39, absorbed the adhesion-inhibitory activity of anti-G9B IgG. This clone contained an insert of approximately 2000 bp and expressed unique 200 and 53 kDa proteins that reacted with monospecific anti-adhesin antibody. The 200 kDa protein also reacted with anti-β-galactosidase IgG, indicating that it is a fusion protein of which 84 kDa represents the streptococcal adhesin. The 84 and 53 kDa proteins are similar in size to the major polypeptides in a streptococcal antigen complex which is associated with the adhesion of G9B to salivacoated hydroxyapatite. The 53 kDa fragment may result from post-translational cleavage of the recombinant polypeptide.