Undergraduate Experimental Biochemistry in Enzymology and Molecular Biology

酶学和分子生物学本科实验生物化学

• 批准号: 8951820
• 负责人: Simon Chang
• 金额: $ 2.88万
• 依托单位: Louisiana State University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-03-01 至 1992-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=8951820&HistoricalAwards=false
• 关键词: Undergraduate; Experimental; Biochemistry; Enzymology; Molecular

This award provides funds to the Department of Biochemistry at Louisiana State University at Baton Rouge to purchase equipment that will help upgrade the core biochemistry lab course. This undergraduate Experimental Biochemistry course will focus on enzymology and molecular biology using a cloned full-length cDNA of Drosophila melanogaster alcohol dehydrogenase (ADH) to provide experimental materials. ADH will be purified from E. coli carrying the ADHcDNA cloned in an expression vector. The purified enzyme will be analyzed to determine the Km values for primary and secondary alcohols and the Ki values for competitive and non- competitive inhibitors. Purified ADH will be reversibly inactivated and reactivated through thio-disulfide interconversion. Effects of Nonsense, Missense and Deletion mutations will be examined by comparing ADH activity and antigenic response in the lysates of E. coli carrying the mutated ADHcDNAs. The ADH insert in M13 phage will be used to teach restriction mapping and DNA sequencing techniques. The principles of nucleic acid chemistry will be illustrated by determining the sequences of di- and tri- ribonucleotides with ribonuclease digestions, chromatography and UV spectra of the products under acidic, neutral and basic conditions. Denaturation of doubly-stranded DNA will be examined. Computer analysis and application are emphasized in appropriate experiments. The grantee is matching with award with non-Federal sources.

该奖项为生物化学系提供资金， 位于巴吞鲁日的路易斯安那州立大学购买设备 这将有助于提升核心生物化学实验课程。 这 本科生实验生物化学课程将侧重于 使用克隆的全长cDNA的酶学和分子生物学 果蝇乙醇脱氢酶（ADH）， 实验材料 从E.大肠杆菌携带 将ADH cDNA克隆到表达载体中。 纯化的酶 将进行分析，以确定初级和 仲醇和Ki值的竞争性和非 竞争性抑制剂 纯化的ADH将可逆地 通过硫代二硫化物相互转化失活和再活化。 无义、错义和缺失突变的影响将是 通过比较ADH活性和抗原反应， E.大肠杆菌中表达突变的ADHcDNA。 ADH插入物 在M13噬菌体中，将用于教导限制性作图和DNA 测序技术。 核酸化学原理 将通过确定二和三的序列来说明。 核糖核苷酸与核糖核酸酶双链反应，色谱法和 酸性、中性和碱性条件下产物的紫外光谱 条件 将检查双链DNA的变性。 计算机分析和应用在适当的强调 实验 该受赠人正在与非联邦奖匹配 源