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通过在单克隆抗体552存在的情况下培养来选择人型支原体PG21缺失突变体。

Selection of Mycoplasma hominis PG21 deletion mutants by cultivation in the presence of monoclonal antibody 552.

作者信息

Jensen L T, Ladefoged S, Birkelund S, Christiansen G

机构信息

Department of Medical Microbiology and Immunology, University of Aarhus, Denmark.

出版信息

Infect Immun. 1995 Sep;63(9):3336-47. doi: 10.1128/iai.63.9.3336-3347.1995.

Abstract

Three mutants of Mycoplasma hominis PG21 were isolated and shown to contain alterations in the size of a repeat-containing gene encoding a surface-localized 135-kDa antigen designated Lmp1. The mutants were isolated by cultivating M. hominis for a 3-month period in the presence of Lmp1-specific monoclonal antibody (MAb) 552. The epitope for MAb 552 was localized at the repeated part of the protein. The gene encoding Lmp1 is part of a transcriptional complex that contains 9.5 direct repeats of 471 bp each. Pure cultures of mutant strains were obtained by subcloning, and three mutants were characterized. The mutants showed deletions of a various number of repeats. The deletions were accompanied by a decrease in size of the proteins. With increasing size of deletions, agglutination and growth inhibition by MAb 552 became less pronounced. Spontaneous aggregation of the mutant M. hominis cells in culture medium was, however, increased, indicating that the repeated elements may be of importance for repulsion of the cells.

摘要

人型支原体PG21的三个突变体被分离出来,结果显示,编码一种定位于表面的135 kDa抗原(命名为Lmp1)的含重复序列基因的大小发生了改变。这些突变体是通过在Lmp1特异性单克隆抗体(MAb)552存在的情况下,将人型支原体培养3个月而分离得到的。MAb 552的表位定位于该蛋白的重复部分。编码Lmp1的基因是一个转录复合体的一部分,该复合体包含9.5个471 bp的正向重复序列。通过亚克隆获得了突变菌株的纯培养物,并对三个突变体进行了表征。这些突变体显示出不同数量重复序列的缺失。这些缺失伴随着蛋白质大小的减小。随着缺失大小的增加,MAb 552引起的凝集和生长抑制作用变得不那么明显。然而,培养基中突变型人型支原体细胞的自发聚集增加,这表明重复元件可能对细胞的排斥很重要。

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