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Negative control gene of streptomyces roseoflavus as well as preparation method and application thereof

A technology of Streptomyces roseus flavum and genes, applied in the biological field, can solve problems such as the prevention and control of cotton Verticillium wilt by undiscovered antibiotics

Active Publication Date: 2012-08-15
HEBEI AGRICULTURAL UNIV.
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there is no report on the use of antibiotics to control cotton verticillium wilt in the current production

Method used

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  • Negative control gene of streptomyces roseoflavus as well as preparation method and application thereof
  • Negative control gene of streptomyces roseoflavus as well as preparation method and application thereof
  • Negative control gene of streptomyces roseoflavus as well as preparation method and application thereof

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Embodiment Construction

[0049] The following examples are used to illustrate the present invention, but are not intended to limit the scope of the present invention. Unless otherwise specified, the raw materials used in the examples are all commercially available, wherein the Streptomyces roseoflavum Men-myco-93-63 was purchased from the Biocontrol Laboratory of Hebei Agricultural University, and Streptomyces coelicolor M145 was purchased from the Institute of Microbiology, Chinese Academy of Sciences ; For the preparation of genomic DNA, see literature: Sambrook J, Russell D W. Molecular cloning: a laboratory manual (3rd Edition). Cold Spring Harbor, New York: Cold spring Harbor Laboratory Press.2001).

[0050] The present invention uses Streptomyces roseoflavum Men-myco-93-63 genomic DNA as a template to perform PCR amplification to obtain a 1121bp fragment, and uses the non-specific primer-anchored PCR (NPA-PCR) method to extend the two ends of the target sequence, The 5' end was extended to 534bp...

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Abstract

The invention relates to a nsdBmgh gene of streptomyces roseoflavus (Men-myco-93-63) as well as a preparation method and application thereof. The preparation method of the nsdBmgh gene of streptomyces roseoflavus comprises the following steps: automatically designing a primer Ns8F and Ns8R through the sequence of a negative control gene nsdB in the metabolic pathway of sky blue streptomycete A3(2) antibiotic, amplifying genomic DNA of the streptomyces roseoflavus (Men-myco-93-63) to obtain fragments of 1121bp, and then extending the two ends of a target sequence by utilizing a nonspecific primer anchoring PCR(NPA-PCR) method so as to obtain the nsdBmgh gene of the streptomyces roseoflavus (Men-myco-93-63). The functions of the nsdBmgh gene are verified through the gene disruption, and the bacteriostatic activity of the gene disruption mutant strain is obviously enhanced as compared with that of metabolin of original strains. The preparation and the application of the nsdBmgh gene provide an important basis for further researching and verifying the synthetic and metabolic pathway and the adjusting and controlling mechanism of the streptomyces roseoflavus, and lay the foundation for the application of high-yield genetic engineering strains of antibiotics for preventing plant diseases.

Description

technical field [0001] The present invention relates to the field of biotechnology, specifically, to Streptomyces roseus flavum ( Streptomyces roseoflavus ) Men-myco-93-63 nsdB mgh Genes, preparation methods and their applications. Background technique [0002] Cotton verticillium wilt, cotton fusarium wilt and cotton bollworm are major pests and diseases in cotton production. With the utilization of insect-resistant cotton and fusarium wilt-resistant varieties, cotton bollworm and cotton bollworm have been effectively controlled. However, cotton verticillium wilt is a worldwide and typical soil-borne disease, and its control is always a worldwide problem. So far, there are no disease-resistant varieties or effective control agents and methods. The disease has spread to dozens of cotton-growing countries and has become the most important threat to cotton production in the world. After the 1990s, the disease continued to break out and become popular in various cotton-pr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/31C07K14/36C12N15/63C12N1/21C12N15/76A01N47/44A01P3/00C12N15/10C12R1/465
Inventor 李亚宁刘大群孟庆芳张立荣李星张娜冀红柳张汀
Owner HEBEI AGRICULTURAL UNIV.
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