DNA Microarray analysis of Clostridium botulinum

描述（由申请人提供）：肉毒梭菌是一种厌氧孢子形成细菌，可产生肉毒神经毒素，肉毒神经毒素是最有效的蛋白毒素，已知可导致人类和动物肉毒中毒。肉毒神经毒素（A ~ G）有7种血清型，由不同的肉毒梭菌株产生.肉毒杆菌，它们在生理学和遗传学上是不同的。神经毒素基因簇包含神经毒素基因和毒素相关蛋白基因，位于不同的C.肉毒杆菌菌株，表明基因簇可能是移动的。PI研究的总体目标是了解神经毒素复合物的生物学以及这些神经毒素在C.肉毒杆菌和相关物种。本研究利用了最近获得的C. A型肉毒杆菌DNA微阵列来自NIH/NIAID赞助的病原体功能基因组学资源中心。我们将使用这些微阵列芯片在全基因组水平上研究神经毒素复合物的产生以及C。肉毒杆菌。在具体目标1中，我们将进行比较基因组微阵列分析，以表征各种C。A型肉毒杆菌菌株，并将它们与未获得毒素基因簇的菌株进行比较。将鉴定产毒素菌株的独特基因并进一步鉴定其特征。在具体目标2中，我们将确定C.肉毒杆菌毒素的产生水平。使用微阵列分析，我们将检查和比较毒素产生野生型菌株和毒素缺陷突变株之间的全局基因转录谱。将分析神经毒素对细胞功能、生长和孢子形成的影响。在拟议的研究中获得的知识将有助于我们了解毒素生产的生物学在C。肉毒杆菌的目的是促进有关神经毒素的知识体系，并开发新的策略，以防止肉毒杆菌中毒。公共卫生相关性：肉毒梭菌产生肉毒神经毒素，这是已知引起人类和动物肉毒中毒的最有效的蛋白质毒素。由于该毒素的效力，该细菌及其神经毒素因其可能用作生物武器而被列为“A类”特选制剂。这些研究有助于进一步了解C.肉毒杆菌的目的是促进有关神经毒素的知识体系，并开发新的策略，以防止肉毒杆菌中毒。

DESCRIPTION (provided by applicant): Clostridium botulinum is an anaerobic spore-forming bacterium that produces botulinum neurotoxin, the most potent protein toxin, which is known to cause botulism in humans and animals. There are seven serotypes of botulinum neurotoxins (A - G), produced from different strains of C. botulinum, that are physiologically and phylogenetically distinct. The neurotoxin gene cluster, containing the genes for the neurotoxin and the toxin associated proteins, is located on different parts of the genome in different C. botulinum strains, indicating that the gene cluster may be mobile. The overall goal of the PI's research is to understand the biology of the neurotoxin complex and the roles of these neurotoxins in the physiology and evolution of C. botulinum and related species. The study proposed here takes advantage of the recently available C. botulinum type A DNA microarray from NIH/NIAID sponsored Pathogen Functional Genomics Resource Center. We will use these microarray chips to examine, at the whole genome level, the role of neurotoxin complex production and the physiology and evolution of C. botulinum. In Specific Aim 1, we will perform comparative genomic microarray analyses to characterize the genomes of various C. botulinum type A strains and compare them to strains that did not acquire the toxin gene cluster. Genes unique to the toxin-producing strains will be identified and further characterized. In Specific Aim 2, we will determine the growth phase-dependent gene expression profiling of C. botulinum in response to the botulinum neurotoxin production levels. Using the microarray analyses, we will examine and compare the global gene transcriptional profiles between toxin-producing wild-type strains and toxin-deficient mutant strains. The impacts of the neurotoxin on cellular functions, growth and sporulation will be analyzed. The knowledge gained in the proposed studies will help us understand the biology of toxin production in C. botulinum with the intent of contributing to the body of knowledge about the neurotoxin and developing novel strategies to prevent botulism. PUBLIC HEALTH RELEVANCE: Clostridium botulinum produces botulinum neurotoxin, which is the most potent protein toxin known to cause botulism in humans and animals. Due to the potency of the toxin, the bacterium and its neurotoxins are classified as the "Category A" Select Agents for their potential use as biological weapons. The proposed studies allow further understanding of the biology of toxin production in C. botulinum with the intent of contributing to the body of knowledge about the neurotoxin and developing novel strategies to prevent botulism.