Saccharopolyspora tuberosa engineering strain with doubled pII gene as well as construction method and application of saccharopolyspora tuberosa engineering strain

A technology of Saccharopolyspora spp. and engineering strains, which is applied in the field of S. saccharopolyspora spp. engineering strains and its construction, and can solve the problems of long fermentation period, low yield of butenyl pleocidins, and many by-products of fermentation broth.

Pending Publication Date: 2022-05-06
HUNAN NORMAL UNIVERSITY
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

However, the wild-type Saccharopolyspora palpitaciens has the problems of long fermentation period, low yield of butenyl pleocidins and many by-products in the ferment

Method used

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  • Saccharopolyspora tuberosa engineering strain with doubled pII gene as well as construction method and application of saccharopolyspora tuberosa engineering strain
  • Saccharopolyspora tuberosa engineering strain with doubled pII gene as well as construction method and application of saccharopolyspora tuberosa engineering strain
  • Saccharopolyspora tuberosa engineering strain with doubled pII gene as well as construction method and application of saccharopolyspora tuberosa engineering strain

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Embodiment Construction

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

[0043] See Table 1 for the relevant descriptions and sequences of the strains, plasmids and primers used in the specific embodiments of the present invention.

[0044] (1) Medium formula and strain culture conditions

[0045] (1) LB medium (100 mL): Mix Yeast extract 0.5 g, Tryptone 1 g and NaCl 1 g, dilute to 100 mL with ultrapure water, adjust the pH value of the solution to 7.0, and add 1.6 g agar powder, sterilized by high-pressure steam at 121°C for 20 min, and set aside;

[0046] (2) CSM Streptomyces Activation Medium (100 mL): mix 4.5 g of TSB, 1 g of Glucose, 0.9 g of Yeast extract and MgSO 4 •7H 2 O 0.22 g, mixed with ultrapure water to 100 mL, 1.6 g of agar powder was added to the solid medium, sterilized by high-pressure steam at 115 °C for 30 min, and set aside;

[0047] (3) TSB medium (100 mL): Tryptic Soy Broth 3 g, dilute to ...

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Abstract

The invention relates to a saccharopolyspora tuberosa engineering strain with doubled pII gene and a construction method and application thereof, the strain is saccharopolyspora tuberosa S.pogona-pII and is preserved in China Center for Type Culture Collection on June 9, 2021, and the preservation number of the strain is CCTCC M 2021706 PII. The construction method comprises the following steps: constructing a pOJ260-PkasO-pII recombinant plasmid on the basis of wild type saccharopolyspora tuberosa, and introducing the recombinant plasmid into the saccharopolyspora tuberosa by utilizing a conjugational transfer method. When the S.pogona-pII is used for synthesizing the butenyl spinosad, the yield of the S.pogona-pII is obviously higher than that of a wild type strain and is 245% of that of the butenyl spinosad of the wild type strain.

Description

technical field [0001] The present invention relates to an engineering strain of Saccharopolyspora mentabilis and its construction method and application, in particular to a strain pII Gene-doubled engineering strain of Saccharopolyspora mentabilis and its construction method and application. Background technique [0002] my country is a large agricultural country, and increasing food production through agricultural development is a major issue related to people's livelihood, but agricultural production is often disturbed by various agricultural diseases and pests. The use of chemical pesticides to reduce agricultural diseases and pests is a common means of farm operations. However, the continuous use of chemical pesticides will cause pests to develop resistance, and the residual chemical pesticides will also cause environmental pollution. [0003] Nitrogen source is a nutrient that is crucial to the growth and development of microorganisms. How to absorb and effectively us...

Claims

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Application Information

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IPC IPC(8): C12N1/21C12N15/74C12N15/66C12N15/31C12P19/62C12R1/01
CPCC07K14/195C12N15/74C12P19/62Y02E50/10
Inventor 夏立秋胡金娟何昊城丁学知
Owner HUNAN NORMAL UNIVERSITY
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