GENETICS OF LAMBDA PROPHAGE - BIOTIN OPERON

Lambda 原噬菌体的遗传学 - 生物素操纵子

• 批准号: 3124421
• 负责人: ALLAN M CAMPBELL
• 金额: $ 26.82万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1978
• 资助国家: 美国
• 起止时间: 1978-06-01 至 1988-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3124421
• 关键词: bacteriophage lambda; prophages; virus genetics

The DNA base sequence of the pI promoter of the lambda-related phage 21 will be determined. We have already sequenced pI from phage 434. Phage 21 is of interest because, unlike 434, it has a different insertion site than lambda. However, its pI promoter responds to the cII protein. Mutations in the pI promoter of lambda will be identified and sequenced. The structural gene for biotin sulfoxide reductase (bisC) will be cloned on a multi-copy plasmid. This should provide us with sufficient bisC protein to obtain a pure product, as well as facilitating genetic analysis. Insertions of Mu Ap 1ac in bisC will be selected so that the regulation, if any, of bisC can be examined. We have found that this enzyme uses the same molybdenum cofactor (product of genes ch1A, ch1B, ch1E, ch1G) as nitrate reductase. The relation of those genes to bisC regulation will be tested. The birA gene, structural gene for the bifunctional protein holoenzyme-synthetase biotin-repressor, already cloned as a 2.1 kb segment, will be sequenced in wild type and mutant form.

Lambda相关噬菌体21的PI启动子的DNA碱基序列将为 决定。 我们已经从噬菌体434测序了Pi。噬菌体21是 兴趣是因为与434不同，它的插入站点与lambda不同。 但是，其PI启动子对CII蛋白有反应。 PI中的突变 Lambda的启动子将被识别和测序。 生物素亚硫氧化物还原酶（BISC）的结构基因将克隆在A上 多拷贝质粒。 这应该为我们提供足够的bisc蛋白 获得纯产品，并促进遗传分析。 插入 将选择BISC中的MU AP 1AC，以便法规（如果有的话）可以 被检查。 我们发现该酶使用相同的钼辅助因子 （基因CH1A，CH1B，CH1E，CH1G的产物）作为硝酸盐还原酶。 关系 将测试那些针对BISC调节的基因。 Bira基因，双功能蛋白的结构基因 全酶 - 合成酶生物素抑制剂（已经被克隆为2.1 kb段）将于 以野生型和突变形式进行测序。