Cellular Responses to Superoxide-Mediated Stress in N. crassa

粗糙猪笼草中细胞对超氧化物介导的应激的反应

• 批准号: 9022177
• 负责人: Donald Natvig
• 金额: $ 12.86万
• 依托单位: University of New Mexico
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-03-01 至 1993-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9022177&HistoricalAwards=false
• 关键词: Cellular; Responses; Superoxide; Mediated; Stress

The goal of the present study is to investigate the nature and magnitude of the genetic response to superoxide-,mediated stress in the fungus Neurospora crassa. The emphasis of the study will be on genes for superoxide dismutase and anonymous genes that are induced in response to superoxide generating agents. Specific goals are to:(1) Continue analysis of the function and regulation of the gene for N. crassa cytosolic CuAn superoxide dismutase (sod-1), which we have recently cloned and sequenced. Recently constructed sod-1 mutants will be studied to determine the importance of CuZn SOD for the aerobic growth and reproduction of N. crassa . Regulation studies will examine the extent to which this gene and the gene (s) for MnSOD (see below) are induced in response to conditions of oxidative stress. In addition, in vitro and in vivo studies will investigate the significance of sequences hypothesized to be important for sod-1 expression. (2) Construct sublcones of the recombinant plasmid carrying the gene for N. crassa MnSOD (currently identified as a 6 Kbp HindIII fragment in a 40 kbp cosmid clone) and create and characterize mutations in this gene using RIP-mediated disruption. These mutant strains, when considered along with sod-1 mutants, will permit genetic and physiological studies of the importance, function and evolution of superoxide dismutases. (3) Perform preliminary experiments toward assessing whether there exists a global superoxide (Sox) response in ascomycetous fungi, similar to that recently proposed for E. coli. We will employ physiological studies of induced resistance to superoxide stress and genetic studies of anonymous superoxide-induced genes in N. crassa., identified by differential screening with labeled cDNAs.

本研究的目的是调查的性质和 对超氧化物介导的应激的遗传反应的大小 真菌粗糙脉孢菌中。 研究的重点将 超氧化物歧化酶基因和匿名基因， 超氧化物生成剂诱导的。 具体 目的是：（1）继续分析功能和调节 N.粗胞质CuAn超氧化物歧化酶 （SOD-1），我们最近克隆和测序。 最近 构建的SOD-1突变体将被研究以确定 铜锌超氧化物歧化酶对好氧生长和繁殖的重要性 n.粗鲁。 监管研究将审查在多大程度上 该基因和MnSOD的基因（见下文）被诱导， 对氧化应激条件的反应。 另外在 体外和体内研究将研究 推测对SOD-1表达重要的序列。（二） 构建携带该基因的重组质粒的亚克隆 奈瑟氏菌的crassa MnSOD（目前鉴定为6 Kbp HindIII 40kbp粘粒克隆中的片段），并产生和表征 使用RIP介导的破坏在该基因中突变。 这些 当沿着考虑SOD-1突变体时， 允许遗传和生理研究的重要性， 超氧化物歧化酶的功能和进化。 (3)执行 初步实验，以评估是否存在一个 全球超氧化物（Sox）的反应，在子囊菌真菌，类似 最近提出的E。杆菌 我们会委聘 超氧胁迫诱导抗性的生理研究 以及N. crassa.，通过用标记的cDNA进行差异筛选来鉴定。