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GENETIC SCREENING METHOD OF NEGATIVE REGULATORY FACTORS OF STREPTOMYCES BIOSYNTHESIS GENE CLUSTER

机译：链霉菌生物合成基因集群负调节因子的遗传筛查方法

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摘要

The present invention provides a screening method of negative regulatory factors of a Streptomyces biosynthesis gene cluster, the method including: constructing a reporter system in a Streptomyces cell, which is mediated by a promoter of a self-owned target gene of the Streptomyces cell, and then randomly mutating Streptomyces with the reporter system by using a random mutation system constructed based on a transposon Himar1; intensively screening Streptomyces strains that have been subjected to random mutation to obtain a Streptomyces strain with high expression of the target gene; performing phage packaging on a genome of the Streptomyces strain with high expression of the target gene and screening out a cosmid with a random insert; and determining the position of the random insert in the genome of the Streptomyces strain with high expression of the target gene by sequencing DNAs of the cosmid.

机译：本发明提供了一种筛选方法的阴性调节因子的筛选方法链霉菌和生物合成基因簇，包括：在介导的链霉菌和细胞中构建报告系统的方法，其由A介导通过使用基于转座子HIMAR1构建的随机突变系统，将链霉菌的自身靶基因的启动子与报告系统随机突变，然后随机突变与报告系统进行随机突变链霉菌。在经受随机突变的温度筛选链霉菌菌株以获得具有高表达的靶基因的链霉菌菌株;在靶基因的高表达和用随机插入筛选靶基因的菌株的基因组上进行噬菌体包装在链霉菌菌株的基因组上，并用随机插入筛分粘接物;并通过测序胶凝体的DNA测定靶基因的高表达，测定随机插入菌株在链霉菌的基因组中的位置。

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公开/公告号US2021079487A1

专利类型
公开/公告日2021-03-18

原文格式PDF
申请/专利权人 ZHEJIANG UNIVERSITY;
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申请/专利号US201917042170
发明设计人 YONGQUAN LI;XUMING MAO;SHUAI LUO;
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申请日2019-03-20
分类号C12Q1/6897;C12N15/76;
国家 US
入库时间 2022-08-24 17:46:56

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