Preliminary Exploration of Molecular Methods for the Specific Detection and Differentiation of the Bdellovibrios

蛭弧菌特异性检测和分化的分子方法初探

• 批准号: 9615515
• 负责人: Henry Williams
• 金额: $ 7.5万
• 依托单位: University of Maryland at Baltimore
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-09-01 至 1998-02-28
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9615515&HistoricalAwards=false
• 关键词: Preliminary; Exploration; Molecular; Methods; Specific

Abstract: 9615515, PI-Williams: Bdellovibrios are procaryotic, parasitic predators which prey on susceptible gram-negative bacilli. Based on the observation that bdellovibrios reduce the population of its prey in laboratory vessels, a logical question is, does the predatory bacterium play some role in the control of microbial populations in nature? Adequate experimentation to resolve this question has not been reported. This is largely due to the limitation in technology to detect and monitor the predator in the environment. The only method currently available for the specific detection of bdellovibrios is the culture technique. As with nearly all cultural methods this technique has substantial limitations. Serological and molecular probes and polymerase chain reaction (PCR) primers which allow for the specific detection of a number of bacterial species have been constructed. However such probes have not been developed for the bdellovibrios. A probe for the bdellovibrios willould allow studies which monitor the numbers of these predators in the environment. The ability to detect those which are parasitic within host cells would allow an estimation of the number of infected cells in the environment and hence define the impact of bdellovibrios on populations of susceptible bacteria. The aim of this researchproposal is to develop a genetic probe and PCR primers which can be used for the specific detection of bdellovibrios in aquatic environments. Initial efforts will be made to identify unique sequences among known nucleic acid sequences for bdellovibrios and to test these as probes and as PCR primers. The objectives and experimental approaches to developing a probe specific for the bdellovibrio group or subgroups of these organisms are described below. 1. Identification and synthesis of specific DNA oligonucleotide sequences for use as detection probes and PCR primers; identification of candidate genomic restriction fragment sequences for use as DNA probes and their production by cloning and/or PCR synthesis; identification of candidate rRNA sequences and construction of rRNA probes. 2. To examine the use and sensitivity of PCR to detect bdellovibrios in laboratory microcosms. 3. To examine the use of genus and species specific probes to confirm the identification of plaques (colonies) of bdellovibrios recovered on culture medium.

摘要：9615515，pi-williams：bdellovibrios是proc虫，寄生虫捕食者，它们捕食易感的革兰氏阴性杆菌。基于Bdellovibrios减少其在实验血管中猎物的种群的观察，一个合乎逻辑的问题是，掠食性细菌在自然界中微生物种群的控制中是否起作用？ 尚未报告足够的实验来解决这个问题。这主要是由于技术限制检测和监测环境中的捕食者。目前可用于特定检测Bdellovibrios的唯一方法是培养技术。与几乎所有文化方法一样，该技术都有很大的局限性。 已经构建了允许对许多细菌物种的特定检测的血清学和分子探针以及聚合酶链反应（PCR）引物。但是，尚未为Bdellovibrios开发此类探针。对Bdellovibrios的探测器将允许研究监测环境中这些捕食者数量的研究。检测宿主细胞中寄生虫的能力将允许估计环境中感染细胞的数量，因此定义了bdellovibrios对易感细菌种群的影响。 这项研究范围的目的是开发一种遗传探针和PCR引物，可用于在水生环境中特定检测Bdellovibrios。最初的努力将在识别已知的bdellovibrios中已知核酸序列之间的独特序列，并将其作为探针和PCR引物测试。 下面描述了针对Bdellovibrio组或这些生物的亚组的特定探针的目标和实验方法。 1。鉴定和合成特定的DNA寡核苷酸序列，用作检测探针和PCR引物；通过克隆和/或PCR合成来鉴定用于用作DNA探针的候选基因组限制片段及其产生；鉴定候选rRNA序列和RRNA探针的构造。 2。检查PCR检测实验室缩影中Bdellovibrios的使用和敏感性。 3。检查使用属和物种特异性探针以确认在培养基上回收的bdellovibrios的斑块（菌落）的鉴定。