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
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[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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