High-yield engineering bacterium of nicomycin x component and its application

A technology of nikkomycin and recombinant engineering bacteria, which is applied in the fields of application, genetic engineering, bacteria, etc., and can solve the problems that the biosynthesis pathway of nikkomycin has not been clearly studied

Inactive Publication Date: 2004-11-24
INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
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Problems solved by technology

[0003] The biosynthetic pathway of ni

Method used

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  • High-yield engineering bacterium of nicomycin x component and its application
  • High-yield engineering bacterium of nicomycin x component and its application
  • High-yield engineering bacterium of nicomycin x component and its application

Examples

Experimental program
Comparison scheme
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Embodiment Construction

[0021] specific implementation plan

[0022] Through the following examples, the present invention will be further described.

[0023] Example 1: Cloning of the sanU gene of Streptomyces chromogenes (CGMCC 4.321) and its function and the construction of a high-yield engineering bacterium of nikkomycin X component

[0024] (1) Cloning of sanU gene

[0025] A, cosmid1, 2, and 3 in the Streptomyces chromogenes (CGMCC 4.321) genomic library were digested with PstI, and the 1.35kb BamHI-ApaI fragment including part of the sanO gene sequence (Nie Liping, Zhang Jihui, Tan Huarong, Acta Microbiology, 41 (1): 59-64, 2001) to perform hybridization, recover the 10.6 kb PstI fragment in cosmid3 according to the positive signal, and then clone it into the PstI site of plasmid M13-.

[0026] B. Digest the 10.6kb PstI fragment with different restriction endonucleases, and select a suitable enzyme to draw a restriction map. And carry out subcloning, sequencing, splicing, use the software F...

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Abstract

The present invention belongs to the field of antibiotic gene engineering, and is especially the process of cloning nicomycin biological synthetic gene sanU from one strain of Streptomyces ansochromogenes 7100 CGMCC 4.321; connecting the synthetic gene sanU with promoter of melanin biosynthetic structure gene, cloning onto the polycloning site of integrative single-copy plamid, transforming wild Streptomyces ansochromogenes 7100 CGMCC 4.321 protoplast to obtain nicomycin X component of the recombinant engineering bacterium in the yield 2 times higher than wild strain. The high-yield engineering bacterium may be used in preparing antiseptic medicines.

Description

technical field [0001] The invention belongs to the field of antibiotic genetic engineering. Specifically, it involves cloning the nikkomycin biosynthesis gene sanU from a Streptomyces ansochromogenes CGMCC 4.321, connecting the sanU gene to the promoter (Pmel) of the melanin biosynthesis structural gene, and then cloning On the multiple cloning site of the single-copy integrated plasmid pSET152, and then transform wild-type Streptomyces chromogenes CGMCC 4.321 protoplasts, the yield of the recombinant engineered strain of nikkomycin X is 2 times higher than that of the wild-type strain about. The high-yield engineered bacteria can be used in the preparation of antibiotic drugs. Background technique [0002] Streptomyces ansochromogenes CGMCC 4.321 is a nikkomycin-producing fungus isolated from soil in Northeast my country. Nicomycin is a class of nucleoside peptide antibiotics, because its structure is similar to the natural substrate UDP-N-acetylglucosamine of chitin sy...

Claims

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Application Information

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IPC IPC(8): C12N1/21C12N15/31C12P21/00
Inventor 谭化荣李毅荣
Owner INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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