Genetic screening method of negative regulatory factors of streptomyces biosynthesis gene cluster
AssigneeZhejiang University
InventorsYongquan LI, Xuming MAO, Shuai LUO
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AssigneeZhejiang University
InventorsYongquan LI, Xuming MAO, Shuai LUO
Estimating the term…
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.
A screening method of negative regulatory factors of a Streptomyces biosynthesis gene cluster, comprising:
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