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Saccharopolyspora pogona engineering strain with tetR26169 gene knockout and application of engineering strain

A technology of tetr26169 and Saccharopolyspora spp. is applied in the field of Saccharopolyspora saccharomyces spp. engineering strains, which can solve the problems of difficulty in genome editing of Saccharopolyspora spp.

Active Publication Date: 2019-07-12
HUNAN NORMAL UNIVERSITY
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  • Abstract
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Problems solved by technology

[0005] At present, there is no report on the knockout of a single gene in Saccharopolyspora. It is very difficult to edit the genome of S. Applications in Saccharopolyspora genome editing are also rarely reported

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  • Saccharopolyspora pogona engineering strain with tetR26169 gene knockout and application of engineering strain
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  • Saccharopolyspora pogona engineering strain with tetR26169 gene knockout and application of engineering strain

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Embodiment Construction

[0028] The present invention will be described in further detail below in conjunction with the accompanying drawings and specific embodiments.

[0029] (1) Specific medium formula and culture conditions

[0030] (1) S. pogona seed activation medium CSM (Complete synthetic medium): Tryptic SoyBroth 4.5g / L, Glucose 0.9g / L, Yeast extract 0.3g / L, MgSO 4 .7H 2 O 0.22g / L. The agar powder that adds 15-20g / L in above-mentioned liquid can be formulated into CSM solid medium. Inoculate the monoclonal or 2% strain preservation solution into a 150mL shake flask filled with 20mL CSM, and incubate at 30°C and 280r / min for 48h;

[0031] (2) S. pogona synthetic fermentation medium SFM (Synthetic fermentation medium): Glucose20.0g / L, Yeast extract 4.0g / L, Tryptone 4.0g / L, KNO 3 1.0g / L, K 2 HPO 4 ·3H 2 O 0.5 g / L, MgSO 4 ·7H 2 O 0.5g / L, FeSO 4 ·7H 2 O 0.01g / L. Inoculate 2% strain activation solution into a 300mL shake flask filled with 50mL SFM, and cultivate for 12 days at 30°C and...

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Abstract

The invention relates to a saccharopolyspora pogona engineering strain with tetR26169 gene knockout and application of the engineering strain. The saccharopolyspora pogona engineering strain with tetR26169 gene knockout is generated in the mode that a transcriptional regulation factor tetR26169 is blocked in a saccharopolyspora pogona genome. The saccharopolyspora pogona engineering strain with tetR26169 gene knockout, namely S. pogona-delta tetR26169 is preserved in China Center for Type Culture Collection on January 22, 2019, and the preservation number of the strain is CCTCC NO:M 2019069. It is expressed through a PCR identification result that the tetR26169 gene in a saccharopolyspora pogona genome is successfully knocked out. It is discovered through thallus OD600 measurement and electron microscope scanning analysis that growth of the engineering strain tetR26169 and the form of the thallus are not influenced. It is proved through a shake flask fermentation result that the butenyl bactericidin yield of the engineering strain saccharopolyspora pogona S. pogona-delta tetR26169 is increased by 2.1 times in comparison with that of an original strain S. pogona.

Description

technical field [0001] The present invention relates to a strain of Saccharopolyspora mensatus tetR26169 gene knockout engineering bacterial strain and application thereof. Background technique [0002] There are various types of agricultural pests in my country. In order to effectively control these major pests such as Lepidoptera, Coleoptera and piercing-sucking mouthparts that seriously threaten my country's agricultural production, chemical pesticides have been relied on for a long time. However, the continuous use of highly toxic and residual chemical pesticides has caused serious environmental pollution and ecological damage, and has seriously affected the safety and export of agricultural products in my country. Therefore, it is particularly urgent to develop high-efficiency biopesticides that are friendly to the ecological environment and safe for humans and animals. [0003] Butenyl spinosyn (butenyl-spinosyn) is a new type of microbial insecticide with contact kil...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/15C12P19/62C12R1/645
CPCC07K14/37C12P19/62
Inventor 何思颖柏丹夏伦万千千夏立秋穰杰何昊城
Owner HUNAN NORMAL UNIVERSITY
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