Molecular identification and functional characterization of fried, a gene require

油炸所需基因的分子鉴定和功能表征

• 批准号: 8142342
• 负责人: JASON Z MORRIS
• 金额: $ 0.65万
• 依托单位: FORDHAM UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-06-01 至 2014-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8142342
• 关键词: Molecular; identification; functional; characterization; fried

DESCRIPTION (provided by applicant): PROJECT ABSTRACT My laboratory's long-term goal is to improve our understanding of growth, metabolism, and cell cycle regulation during Drosophila development. This proposal focuses on the molecular identification and functional characterization of fried. The three fried mutants were identified in the same EMS clonal screen that identified mutants in gatA, the gene that we characterized in meeting the specific aims of NIH R15 grant GM074735-01. As with gatA, mutant clones of fried in the germ cells of the ovary result in chromosome morphology defects in the nurse cells and oocyte followed by egg chamber arrest and degradation (Morris, Navarro et al. 2003). In addition, both mutants show growth defects and developmental arrest, though the fried larvae are more variable in size than gatA, and fried mutants arrest development post-pupariation while gatA mutants arrest as larvae (Morris, Bergman et al. 2008) Study of fried is highly significant because understanding fried will likely shed light on control of growth and cell cycle regulation. The initiatives proposed in this grant are additionally significant because they will greatly enhance the research program at Fordham College at Lincoln Center and will enable the training of 6-10 undergraduates in genetics and cell biology research. No other lab is studying fried, so the work proposed here is very unlikely to be performed elsewhere. Specific Aim 1: To characterize the growth and arrest phenotypes and to describe cell cycle defects in the fried EMS alleles. Specific Aim 2: To identify the gene defined by the mutant fried alleles by candidate gene sequencing and transgenic rescue, to molecularly and genetically characterize these alleles, and to determine the fried expression pattern by RTPCR and in situ hybridization. Specific Aim 3: To analyze the function of fried by using Western Blotting and Immunohistochemistry to expression and activation of growth regulators in fried mutants and by using genetic assays to determine which pathways interact with fried mutations. PUBLIC HEALTH RELEVANCE: PROJECT NARRATIVE Drosophila lacking fried gene function grow poorly and arrest development. Appropriate regulation of growth is one of the most fundamental aspects of development, and many regulators of growth in Drosophila also have essential functions in humans. We propose to identify which gene is disrupted in fried mutants and to determine the function of fried in wild type Drosophila.

描述（由申请人提供）： 我实验室的长期目标是提高我们对果蝇发育过程中生长、代谢和细胞周期调控的理解。本研究主要针对油炸食品的分子鉴定和功能特性进行研究。在鉴定gatA突变体的相同EMS克隆筛选中鉴定了三种油炸突变体，gatA是我们表征为满足NIH R15授权GM 074735 -01的特定目标的基因。与加塔一样，卵巢生殖细胞中fried的突变克隆导致滋养细胞和卵母细胞中的染色体形态缺陷，随后是卵室停滞和降解（Morris，Navarro et al. 2003）。此外，两种突变体都显示出生长缺陷和发育停滞，尽管油炸的幼虫在大小上比加塔更可变，并且油炸的突变体在蛹化后停滞发育，而加塔突变体在幼虫时停滞（Morris，Bergman等人，2008）油炸的研究是非常重要的，因为理解油炸可能会阐明生长和细胞周期调控的控制。在这项赠款中提出的倡议是额外的重要，因为它们将大大提高林肯中心的福德姆学院的研究计划，并将使6-10名本科生的遗传学和细胞生物学研究的培训。没有其他实验室正在研究油炸，所以这里提出的工作不太可能在其他地方进行。具体目标1：描述生长和停滞表型，并描述油炸EMS等位基因中的细胞周期缺陷。具体目标二：通过候选基因测序和转基因拯救鉴定突变fried等位基因所定义的基因，对这些等位基因进行分子和遗传学表征，并通过RTPCR和原位杂交确定fried表达模式。具体目标3：通过Western Blotting和免疫组化技术分析fried突变体中生长调节因子的表达和激活情况，并通过遗传学分析确定哪些途径与fried突变相互作用，从而分析fried的功能。 公共卫生相关性： 项目叙述缺乏油炸基因功能的果蝇生长不良，发育停滞。适当的生长调节是发育的最基本方面之一，果蝇中的许多生长调节因子在人类中也具有重要功能。我们建议，以确定哪些基因被破坏的油炸突变体，并确定野生型果蝇油炸的功能。