Improvement of genetic analysis in the pathogenic zygomycete Mucor circinelloides

 描述（由申请人提供）：该提案旨在开发在放线菌真菌卷枝毛霉中增强的遗传分析系统。M.卷枝霉是一种引起毛霉病的机会性真菌病原体。毛霉菌病的发病率正在增加，由于越来越多的免疫功能低下的个人。M. circinelloides也作为一个模型系统，研究光传感和理解RNAi介导的基因表达调控。此外，M. circinelloides是生物燃料生产的候选者。然而，由于1）用于转化的标记数量有限; 2）转化效率低和同源基因替换频率低;和3）缺乏进行遗传分离分析的能力，该真菌系统的遗传分析是有限的。在本文中提出的研究中，我们的目标是通过提出三个目标来提高我们在这个重要的真菌系统中进行遗传分析的能力：1）构建可回收的pyrG标记系统; 2）增强基因破坏中的同源重组;以及3）在M.卷枝藻

 DESCRIPTION (provided by applicant): This proposal aims to develop enhanced genetic analysis systems in the zygomycete fungus Mucor circinelloides. M. circinelloides is an opportunistic fungal pathogen that causes mucormycosis. The incidence of mucormycosis is increasing due to a rising number of immunocompromised individuals. M. circinelloides also serves as a model system to study light sensing and understand RNAi-mediated gene expression regulation. In addition, M. circinelloides is a candidate for biofuel production. However, the genetic analysis of this fungal system is limited due to 1) a limited number of markers for transformation; 2) inefficient transformation and low frequent homologous gene replacement; and 3) lack of ability to perform genetic segregation analysis. In the studies proposed within this document, we aim to enhance our ability for genetic analysis in this important fungal system by proposing three aims: 1) construction of a recyclable pyrG marker system; 2) enhancement of homologous recombination in gene disruptions; and 3) a pilot approach for mating and genetic analysis in M. circinelloides.