HOST TEMPERATURE GROWTH OF C NEOFORMANS

新型隐球菌引起危及生命的脑膜脑炎 在免疫抑制和正常宿主中。尽管治疗与 抗生素B和三唑类，这种感染的管理仍然是 由于缺乏杀真菌方案和发展， 抗药性的证据。这项提案的基础是基因控制 对于新型隐球菌的宿主温度生长，是基于 假设C.新形式适应37摄氏度的宿主 通过表达或抑制某些基因， 在这种环境压力下的生存和成长。的这种圣贤观 研究环境压力，如温度调节毒性， 潜力和微生物已经非常有效地用于 病原菌优雅地阐明分子机制， 疾病最近，我们证明了C.新生菌， 阐明疾病的分子机制最近我们 证明了C. neoformans可以类似地作为模型系统， 战略例如，我们发现C. neoformans使用 钙调神经磷酸酶A（CNAl）基因在信号通路中的37度 摄氏度、pH、pCO 2生长和毒力。CNAl是第一个 在C.新生儿及其 重要性不能被经典的模式真菌预测， 作为S.啤酒。沿着先前的基因数据和动物研究 很明显，温度生长表型对此至关重要， 从这些研究中得到的病原体和概念， 其他真菌病原体。在本提案中，我们将使用以下内容 策略：1）插入突变，2）调控启动子陷阱，3） cDNA文库消减，和4）差异显示RT-PCR以捕获 一系列高温调节基因和/或必需基因 在37摄氏度下生长。我们的焦点将使用这些策略 然后我们将分离，测序和验证 通过在动物模型中敲除突变体来研究这些基因的重要性。的 直接的表型研究使我们能够专注于杀真菌靶点（即， 无法在37摄氏度下生存）我们的梳理策略 梭用于高温生长基因的新生儿基因组可以 代表了新的抗真菌靶点验证的潜力 毒品

Cryptococcus neoformans produces a life-threatening meningoencephalitis in both immunosuppressed and normal hosts. Despite treatments with amphotericin B and triazoles, the management of this infection remains suboptimal because of the lack of fungicidal regimens and development of drug resistance. The foundation for this proposal, Genetic controls for host temperature growth of Cryptococcus neoformans, is based on the hypothesis that C. neoformans adapt to 37 degrees Celsius in the host by expressing or repressing certain genes which are essential for its survival and growth under this environmental stress. This concept of studying environmental stresses such as temperature regulating virulence potential and the microorganism has been very effectively used in pathogenic bacteria to elegantly elucidate molecular mechanisms of disease. Recently, we have proven that C. neoformans bacteria to elegantly elucidate molecular mechanisms of disease. Recently, we have proven that C. neoformans can similarly act as a model system for this strategy. For instance, we discovered that C. neoformans uses the calcineurin A (CNAl) gene in the signaling pathway for 37 degrees Celsius, pH, pCO2 growth and virulence. CNAl is the first gene identified with elevated temperature growth in C. neoformans and its importance could not have been predicted by classical model fungi such as S. cerevisiae. Along with previous genetic data and animal studies it is clear that the temperature growth phenotype is essential to this pathogen and concepts from these studies can probably by extrapolated to other fungal pathogens. In this proposal, we will use the following strategies: 1) insertional mutagenesis, 2) regulated promoter trap, 3) cDNA library subtraction, and 4) differential display RT-PCR to capture a series of elevated-temperature regulated genes and/or essential genes for growth at 37 degrees Celsius. Our focus will use these strategies for identification but then we will isolate, sequence, and validate the importance of these genes by knockout mutants in animal models. The direct phenotype study allows us to focus on fungicidal targets (i.e. inability to survive at 37 degrees Celsius) Our strategies of combing the C. neoformans genome for elevated temperature growth genes can represent the potential for validation of antifungal targets for new drugs.