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Saccharopolyspora spinosa recombinant strain with double bldD genes

A technology of Saccharopolyspora spinosa and recombinant strains, applied in microorganism-based methods, fungi, microorganisms, etc., can solve problems such as integration difficulties and low efficiency, and achieve biosynthesis promotion, increase copy number, and good genetic stability. Effect

Inactive Publication Date: 2015-10-21
HUNAN NORMAL UNIVERSITY
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  • Abstract
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AI Technical Summary

Problems solved by technology

[0004] At present, the genetic modification of S. spinosa mainly adopts the methods of protoplast transformation and conjugative transfer, but the efficiency of both is very low, and there are certain difficulties in integrating foreign genes into the genome of S. spinosa.

Method used

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  • Saccharopolyspora spinosa recombinant strain with double bldD genes
  • Saccharopolyspora spinosa recombinant strain with double bldD genes
  • Saccharopolyspora spinosa recombinant strain with double bldD genes

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Embodiment Construction

[0042] The present invention will be described in further detail below in conjunction with the accompanying drawings and specific embodiments.

[0043] In this embodiment bld Gene-doubled Saccharopolyspora spinosa recombinant strain, said bld Gene-doubled recombinant strain of Saccharopolyspora spinosa consisting of the bald gene bld Generated by integration into the S. spinosa genome.

[0044] In this embodiment bld Gene-doubled recombinant strain of S. spinosa, S. spinosa S. spinosa-BldD (Saccharopolyspora spinosa S. spinosa-BldD ) was deposited in China Center for Type Culture Collection (CCTCC for short, address: Wuhan University, Wuhan, China) on May 31, 2015, and the strain preservation number is CCTCC NO: M 2015347.

[0045] (1) The bald gene bldD, Cloned from Saccharopolyspora spinosa, its sequence is as follows:

[0046] atgggcgact acgccaaggc gctgggcagc aagctccgcg ctatccgcca gcagcagggt 60

[0047] ctgtcgctgca cggcgtcgag cagaagtctg gcgggcggtg gaaggccgtg gtcg...

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Abstract

The invention discloses a saccharopolyspora spinosa recombinant strain with double bldD genes. The provided saccharopolyspora spinosa recombinant strain with double bldD genes is prepared by integrating bldD gene into saccharopolyspora spinosa genome. The provided saccharopolyspora spinosa recombinant strain with double bldD genes is named as S.spinosa-BldD, and is preserved in the China Center for Type Culture Collection (CCTCC) in May 31st, 2015, and the preservation number is CCTCC No. M2015347. The PCR detection results show that bldD gene has been successfully integrated into a target bacterial strain chromosome. In the plate culture, people find that in BHI and TSB plates, the spores of recombinant strains are prominently inhibited. The results of shaking flask fermentation show that the spinosad yield of recombinant strain is increased by 1.35 times compared with that of a referential strain.

Description

technical field [0001] The present invention relates to gene recombination, in particular to increasing bld Gene copy number, obtain spinosad high-yielding strain S. spinosa-BldD . Background technique [0002] Environmental protection and food safety issues are the focus of attention in today's society. The use of residual chemical pesticides leads to the pollution of agricultural products and the destruction of the ecological environment. The negative impact on human beings cannot be ignored. The development of green and environment-friendly biopesticides is an inevitable trend in the development of the pesticide industry. Spinosad, a secondary metabolite of Saccharopolyspora spinosa, has a broad insecticidal spectrum and a unique insecticidal mechanism. It has excellent killing effects on various crop pests such as Coleoptera, Lepidoptera, Homoptera, and Thysanoptera. Its characteristics of safety, rapidity and low residue make it one of the most promising new biopest...

Claims

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Application Information

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IPC IPC(8): C12N1/15C12P19/62C12R1/645
Inventor 夏立秋蔡妹丁学知孙运军
Owner HUNAN NORMAL UNIVERSITY
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