REGULATION OF EXPRESSION OF BORRELIA BURGDORFERI BMPC

伯氏疏螺旋体BMPC的表达调控

• 批准号: 2887734
• 负责人: Felipe Cardenas Cabello
• 金额: $ 28.6万
• 依托单位: NEW YORK MEDICAL COLLEGE
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-07-01 至 2002-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2887734
• 关键词: Borrelia; Escherichia coli; SCID mouse; bacterial cytopathogenic effect; bacterial genetics; bacterial proteins; gene expression; genetic regulation; immunoelectron microscopy; northern blottings; protein localization; protein purification; transfection

Lyme disease is a multisystem, tick-borne chronic infection caused by the persistence of the spirochete Borrelia burgdorferi. B. burgdorferi has the ability to proliferate in a variety of niches in reservoir and human hosts as well as in the vector lxodes scapularis. Preliminary evidence indicates that differential gene expression resulting from environmental stimuli in one of the mechanisms that allows bacterial to adapt rapidly to fast changing environments found during infection and disease of vectors and hosts. We and others have described a family of chromosomal genes (bmpA-D) encoding homologous 36.9-39.8 kDa lipoproteins of the p39 family. The tandem chromosomal location of the bmp genes, their homology and overlapping regulatory signals together with the potential surface location of the proteins they encode in addition to their apparent up regulation by environmental stimuli suggests the hypothesis that these proteins may be important for B. burgdorferi survival in different environments. In this project, we propose to study the effects environmental stimuli have on bmpC, a member of this gene family first described by us, both in vitro and in vivo in mice and ticks, in an effort to improve our understanding of the mechanisms employed by B. burgdorferi to enable it to persist and thrive in vectors and hosts. The long-range goal of our efforts is to improve understanding of B. burgdorferi gene expression despite the lack of genetic systems that would facilitate this work. The specific aims of the current project are: 1) identify anti-BmpC Mab and polyclonal antibodies not cross-reactive with BmpD, BmpA and BmpB and use them to characterize BmpC protein and confirm its localization and lipidation in B. burgdorferi 297; 2) Determine levels of expression and mechanisms of regulation of bmpC under different in vitro and in vivo conditions; and 3) Determine genetic organization and DNA structural and regulatory sequences of bmpC in B. burgdorferi, B. afzelii and B. garinii genospecies and in B. burgdorferi 297 grown under different conditions in vitro and in vivo. The latter studies will indicate whether different levels of transcription are secondary to changes in the regulatory and structural DNA sequences of the bmpC gene.

莱姆病是一种多系统、蜱传的慢性感染， 伯氏疏螺旋体的持续存在B。burgdorferi 有能力在水库的各种小生境中繁殖， 人宿主以及在载体肩突硬蜱中。 初步 有证据表明，差异基因表达导致的 环境刺激是允许细菌 迅速适应感染期间快速变化的环境， 病媒和宿主的疾病。 我们和其他人描述了一个家庭， 染色体基因（bmpA-D）编码同源36.9-39.8 kDa p39家族的脂蛋白。 串联染色体的位置， BMP基因，它们的同源性和重叠的调控信号一起 它们编码的蛋白质的潜在表面位置 除了受环境刺激的明显上调外， 提出了这些蛋白质可能对B很重要的假设。 Burgdorferi生存在不同的环境中。 本课题 建议研究环境刺激对bmpC的影响， 这个基因家族的成员，首先由我们描述，无论是在体外和在 在小鼠和蜱虫体内，努力提高我们对 B采用的机制。burgdorferi使其能够持续和蓬勃发展 在载体和宿主中。 我们努力的长期目标是改善 理解B。尽管缺乏Burgdorferi基因表达， 基因系统，这将有助于这项工作。 本项目的具体目标是：1）鉴定抗BmpC单克隆抗体 以及不与BmpD、BmpA和BmpB交叉反应的多克隆抗体 并利用它们来表征BmpC蛋白并确定其定位 和在B中的脂化。burgdorferi 297; 2）测定表达水平 并探讨了不同体外和体内条件下bmpC的调控机制。 体内条件;和3）确定遗传组织和DNA B中bmpC结构和调控序列。burgdorferi，B. afzelii和B. garinii基因种和B. burgdorferi 297生长在 在体外和体内的不同条件下。 后一项研究将 表明不同转录水平是否继发于 bmpC基因的调控和结构DNA序列的变化。