MOLECULAR STUDIES OF T PALLIDUM TPRJ, TPRG, AND TPRE GENES

T PALLIDUM TPRJ、TPRG 和 TPRE 基因的分子研究

• 批准号: 6347208
• 负责人: LOLA Virginia STAMM
• 金额: $ 15.71万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-09-01 至 2001-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6347208
• 关键词: Treponema pallidum; active immunization; antibacterial antibody; antibody formation; antigen antibody reaction; bacterial antigens; bacterial genetics; bacterial proteins; clinical research; cooperative study; disease /disorder model; gene expression; genetic polymorphism; human subject; laboratory rabbit; nucleic acid sequence; syphilis

Treponema pallidum is the etiologic agent of syphilis, a multistage sexually transmitted disease that is a global health problem. Identification of virulence determinants and protective immunogens of T. pallidum has been hindered by the non-cultivable nature of this spirochete. Although T. pallidum has a cellular architecture similar to Gram negative bacteria, its outer membrane contains rare, poorly immunogenic proteins whose identified and functions remain elusive. Using TnphoA mutagenesis, we identified a gene (tprJ) encoding a protein with homology to the major surface protein of T. denticola, a periodontal pathogen. The tprJ is a member of a polymorphic multi-gene family encoding 12 proteins (TprA- L), that are divided into three subgroups. Recent studies have shown heterogeneity in the subgroup 3 (tprJ, tprE, and tprG) genes. We hypothesize that functional activities such as attachment, invasion, etc. The long-term goal of our studies is to develop a better understanding of variation of the subgroup 3 tpr genes and their corresponding proteins. The specific aims for the project are: (1) To detect nucleotide sequence variations within or flanking the tprJ, tprE, and tprG genes of T. pallidum obtained a various timepoints from chances of intradermally infected rabbits; (2) To examine expression of the TprJ, TprE and TprG proteins in T. pallidum and to assess the kinetics of the rabbit IgG antibodies to the TprJ, TprE and TprG proteins and to peptides representing the variable and/or conserved regions of the Tpr proteins; (4) To determine if immunization of rabbits with the TprJ, TprE, and TprG proteins of peptide protects against infection with T. pallidum and/or selects for phenotypic variants and (5) To evaluate nucleotide sequence variability of the tprJ, tprE, and tprG genes of T. pallidum in clinical specimens from syphilis patients attending local STD clinics.

梅毒螺旋体是梅毒的病原体，梅毒是一种多阶段性传播疾病，是一个全球性的健康问题。梅毒螺旋体的不可培养性质阻碍了梅毒螺旋体毒力决定簇和保护性免疫原的鉴定。尽管梅毒螺旋体具有与革兰氏阴性细菌相似的细胞结构，但其外膜含有罕见的、免疫原性差的蛋白质，其鉴定和功能仍然难以捉摸。利用 TnphoA 诱变，我们鉴定了一个基因 (tprJ)，其编码的蛋白质与牙周病原体 T. denticola 的主要表面蛋白质具有同源性。 tprJ 是编码 12 种蛋白质 (TprA-L) 的多态性多基因家族的成员，这些蛋白质分为三个亚组。最近的研究表明亚组 3（tprJ、tprE 和 tprG）基因存在异质性。我们假设附着、入侵等功能活动。我们研究的长期目标是更好地了解亚组 3 tpr 基因及其相应蛋白质的变异。该项目的具体目标是：（1）检测从皮内感染兔子的机会获得的不同时间点的梅毒螺旋体的tprJ、tprE和tprG基因内部或侧翼的核苷酸序列变异； (2) 检查梅毒螺旋体中 TprJ、TprE 和 TprG 蛋白的表达，并评估兔 IgG 抗体对 TprJ、TprE 和 TprG 蛋白以及代表 Tpr 可变和/或保守区的肽的动力学蛋白质； (4) 确定用肽的 TprJ、TprE 和 TprG 蛋白免疫兔子是否可以防止梅毒螺旋体感染和/或选择表型变异，以及 (5) 评估 tprJ、tprE 和 TprG 的核苷酸序列变异性当地 STD 诊所就诊的梅毒患者临床标本中梅毒螺旋体的 tprG 基因。