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Recombinant streptomyces lydicus with cellulase activity and construction method and application thereof

A technology of Streptomyces lyddi and cellulase, applied in the field of recombinant Streptomyces lyddi and its construction, can solve the problems of single antibacterial mechanism, narrow antibacterial spectrum, and limited types of metabolites

Active Publication Date: 2014-03-12
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, as a natural strain, the metabolites of a single strain are limited, and there will be a single antibacterial mechanism, poor antibacterial ability, and narrow antibacterial spectrum. When two or more active substances act on pathogenic bacteria together, a synergistic effect can be produced. , very good at inhibiting the growth of pathogenic bacteria

Method used

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  • Recombinant streptomyces lydicus with cellulase activity and construction method and application thereof
  • Recombinant streptomyces lydicus with cellulase activity and construction method and application thereof
  • Recombinant streptomyces lydicus with cellulase activity and construction method and application thereof

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Experimental program
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Effect test

Embodiment 1

[0046] Embodiment 1, construct the recombinant Streptomyces lydidis of anti-phytopathogenic fungus

[0047] 1. Construction of cellulase gene expression vector pIB139-glu

[0048] The name of the cellulase gene expression vector constructed in this example is pIB139-glu, which contains the cellulase gene (the coding sequence is the 14th-1495th position of SEQ ID No. 2, encoding the protein of SEQ ID No. 1). The promoter in pIB139-glu that initiates the transcription of the cellulase gene is the erythromycin resistance gene promoter (SEQ ID No. 3).

[0049] The specific construction method of pIB139-glu is as follows: the cellulase gene fragment with NdeI and EcoRI restriction sites at both ends shown in SEQIDNo. pIB139 (the erythromycin resistance gene promoter PermE carrying SEQIDNo.3) was ligated to obtain a recombinant expression vector pIB139-glu in which the segment between NdeI and EcoRI of pIB139 was replaced with the cellulase gene.

[0050] Escherichia coli DH5α was...

Embodiment 2

[0064] Example 2. The antibacterial activity of Streptomyceslydicus AG02 is higher than that of Streptomyceslydicus A02CGMCCNo.1654

[0065] In this embodiment, the culture medium used is as follows:

[0066] Gao's No. 1 slant medium: K 2 HPO 4 0.5g, NaCl0.5g, KNO 3 1.0g, FeSO 4 ·7H 2 O0.01g, MgSO 4 ·7H 2 O0.5g, soluble starch 20g, agar 20g, water 1000ml; pH7.2-7.4.

[0067] Seed medium: 15g soybean grains (boil with distilled water for 0.5-1h, take the filtrate), 5g peptone, 2.5g ammonium sulfate, 20g glucose, 10g starch, 0.25g magnesium sulfate, 0.2g potassium dihydrogen phosphate, 5g sodium chloride, Make an aqueous solution, adjust the pH to 7-8, add 1g of calcium carbonate, and add water to make it 1000ml.

[0068] Fermentation medium: 15g soybean grains (boil with distilled water for 0.5-1h, take the filtrate), 5g peptone, 2.5g ammonium sulfate, 10g sucrose, 10g starch, 0.25g magnesium sulfate, 0.2g potassium dihydrogen phosphate, 5g sodium chloride, Make an aqu...

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Abstract

The invention discloses a recombinant streptomyces lydicus with a cellulase activity and a construction method and an application of the recombinant streptomyces lydicus. The method for constructing the recombinant streptomyces lydicus with the cellulase activity comprises the following steps of: leading encoding genes of cellulose shown by SEQ ID No.1 to the streptomyces lydicus as host bacteria; and screening to obtain the recombinant streptomyces lydicus with the cellulase activity. The recombinant streptomyces lydicus with the cellulase activity constructed by the invention enhances the antimicrobial activity of the streptomyces lydicus as the host to plant pathogenic fungi, particularly, the antimicrobial activity to watermelon blast fungus is twice as much as that of the host streptomyces lydicus.

Description

technical field [0001] The invention relates to a genetically engineered bacterium and its construction method and application, in particular to a recombinant streptomyces lydidi with cellulase activity and its construction method and application. Background technique [0002] my country is a large agricultural country based on planting. Many major diseases, such as gray mold, anthracnose, tomato leaf mold, watermelon wilt, wheat head blight, etc., have caused serious losses to agricultural production. The economic loss can be reduced by about 30 billion yuan through the use of pesticides every year. However, the amount of chemical pesticides used accounts for more than 80% of the amount of pesticides used, causing serious environmental pollution, pesticide residues, increased resistance, etc., which seriously threaten human health and the ecological environment. . Therefore, biological control is an inevitable trend in the development of pesticides. Microorganisms and the...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/76C12N1/21A01N63/00A01N63/02A01P3/00C12R1/465
Inventor 吴慧玲刘伟成刘建华董丹卢彩鸽刘霆张殿朋张涛涛田兆丰卢向阳
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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