Cloning, Structure and Regulation of the sp56 Gene

sp56基因的克隆、结构和调控

• 批准号: 9509863
• 负责人: Jeffrey Bleil
• 金额: $ 10万
• 依托单位: The Scripps Research Institute
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-09-01 至 1996-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9509863&HistoricalAwards=false
• 关键词: Cloning; Structure; Regulation; sp56; Gene

9509863 Bleil The primary interest of Dr. Bleil's laboratory is analysis of mammalian fertilization at the molecular level, combining techniques of cell biology, biochemistry and molecular genetics. He has identified a mouse sperm protein, a lectin called sp56, which has many properties expected of the protein responsible for sperm recognition of the mouse egg's zona pellucida. He has recently isolated and cloned the cDNA encoding sp56. Amino acid sequence derived from this cDNA indicates that: 1) the protein is translated on the endoplasmic reticulum, 2) has no transmembrane domain, and 3) is a member of a superfamily of extracellular protein receptors. Probes directed against sp56 and its mRNA reveal that transcription of the sp56 gene and translation of its mRNA occur exclusively in spermatogenic cells of the mouse. The long term goals to be addressed by studies described here are: 1) to genetically prove or disprove the hypothesis that sp56 is the sperm protein which recognizes the mouse egg zona pellucida, mediating sperm binding to the egg's zona pellucida, and 2) to determine the molecular structure of the sp56/ZP3 complex. In order to attain these goals, the gene encoding sp56 must be identified and isolated. The specific aims of the proposed study are as follows: 1) To use cDNA encoding sp56, currently available in this laboratory, to screen a mouse genomic library. Genomic DNA clones, including sequence upstream of the translated region, will be purified and sequenced; 2) To analyze the sequence of the sp56 gene in order to identify regulatory sequences which might encode specific expression in testes. Comparison with other, previously identified genes which are expressed in testes may reveal common regulatory sequences. ***

9509863 Bleil Bleil 博士实验室的主要兴趣是在分子水平上分析哺乳动物受精，结合细胞生物学、生物化学和分子遗传学技术。他发现了一种小鼠精子蛋白，一种名为 sp56 的凝集素，它具有许多负责精子识别小鼠卵子透明带的蛋白质所具有的特性。他最近分离并克隆了编码sp56的cDNA。源自该 cDNA 的氨基酸序列表明：1) 该蛋白质在内质网上翻译，2) 没有跨膜结构域，3) 是细胞外蛋白质受体超家族的成员。 针对 sp56 及其 mRNA 的探针揭示了 sp56 基因的转录及其 mRNA 的翻译仅发生在小鼠的生精细胞中。这里描述的研究要解决的长期目标是：1）从遗传学上证明或反驳这样的假设：sp56是识别小鼠卵子透明带的精子蛋白，介导精子与卵子透明带的结合，2）确定sp56/ZP3复合物的分子结构。为了实现这些目标，必须鉴定并分离编码 sp56 的基因。本研究的具体目的如下： 1）利用本实验室现有的编码sp56的cDNA，筛选小鼠基因组文库。基因组 DNA 克隆，包括翻译区域上游的序列，将被纯化和测序； 2)分析sp56基因的序列，以确定可能编码睾丸中特异性表达的调控序列。与先前鉴定的在睾丸中表达的其他基因的比较可能揭示共同的调控序列。 ***