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Marine actinomycete genome library and construction method thereof

A technology of genome library and marine actinomycetes, applied in the field of genome library construction, can solve the problem of lack of gene clusters of natural products and other problems

Inactive Publication Date: 2017-05-10
FOSHAN UNIVERSITY
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Problems solved by technology

[0004] In view of the above deficiencies in the prior art, the object of the present invention is to provide a marine actinomycete genome library and its construction method, aiming at solving the lack of gene clusters of natural products in the prior art allogeneic expression strain S. arenicola CNS-205 The problem

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  • Marine actinomycete genome library and construction method thereof

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Embodiment 1

[0046] 1. Materials and reagents

[0047] Materials: marine actinomycete S. arenicola CNS-205 is preserved by the author's laboratory;

[0048] Reagents: Bam HI restriction endonuclease and plasmid mini-extraction kit were purchased from TaKaRa Company; proteinase K and lysozyme were purchased from Sigma Company; pWEB TM Cosmid Cloning Kit was purchased from Epicentre; the rest were domestic analytical reagents.

[0049] 2. Main instruments

[0050] Low-temperature ultracentrifuge 5415R (Eppendorf Company); constant temperature and humidity incubator PYX-150H-B (Keli Instruments); constant temperature circulator HX-2015 (Zhengzhou Changcheng Technology Industry and Trade Co., Ltd.); constant temperature culture shaker ZHWY-211B ( Shanghai Zhicheng Analytical Instrument Manufacturing Co., Ltd.); gel imaging analyzer (BIO-RAD company Gel Documentation 2000); ND-2000 micro-UV spectrophotometer (Beijing Tuo Morgan Biotechnology Co., Ltd.); Powerpac Basic electrophoresis instrum...

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Abstract

The invention discloses a marine actinomycete genome library and a construction method thereof. The construction method includes: extracting DNA of marine actinomycete S.arenicola CNS-205; adopting a syringe needle of an injector to randomly shear the extracted DNA; adopting terminal repairase to repair the randomly shorn DNA, and connecting with a carrier; adopting packaging protein to package into bacteriophage, and infecting a host cell to obtain the genome library of the marine actinomycete S.arenicola CNS-205. The invention aims to construct the genome library by taking the marine actinomycete S.arenicola CNS-205 which is rare for the convenience of heterologously expressing a secondary metabolism product having biological activity in the strain to biologically synthesize related gene clusters and analyzing genetic functions of the gene clusters so to apply metabolic engineering or combinatorial biosynthesis methods in compound structure modifying.

Description

technical field [0001] The invention relates to the field of genome library construction, in particular to a marine actinomycete genome library and a construction method thereof. Background technique [0002] Actinomycetes are an important source of bioactive substances, especially natural medicines. Facing the increasingly severe drug resistance of pathogenic microorganisms and a large number of repeated discoveries in the screening process of natural medicines, how to use new technologies, new resources, and new models to screen and discover new active secondary metabolites has become an urgent research in the development of natural medicines. important topic. However, marine actinomycetes may adjust their secondary metabolic processes under the selective pressure of the special marine environment (high salt, high pressure, low temperature, low oxygen, low light, and oligotrophic), thereby producing and accumulating bacteria with special chemical structures and biological...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10C40B40/08C40B50/06
CPCC12N15/1093C40B40/08C40B50/06
Inventor 马艳玲陈锦灵刘富来黄桂东曾荣
Owner FOSHAN UNIVERSITY
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