Negative regulator gene of streptomyces roseofulvus as well as preparation method and application thereof

A Streptomyces roseus flavum, genetic technology, applied in the biological field, can solve the problems of easy resistance of pathogens, long-term use of chemical agents, and large dosage

Inactive Publication Date: 2011-10-12
HEBEI AGRICULTURAL UNIV.
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  • Abstract
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  • Claims
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Problems solved by technology

At present, fungicides such as triazoles and pyrimidine amines are mainly used in production to control powdery mildew and gray mold. However, the use of chemical agents take

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  • Negative regulator gene of streptomyces roseofulvus as well as preparation method and application thereof
  • Negative regulator gene of streptomyces roseofulvus as well as preparation method and application thereof
  • Negative regulator gene of streptomyces roseofulvus as well as preparation method and application thereof

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Embodiment Construction

[0039]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).

[0040] 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 an nsdBmgh gene of Streptomyces roseoflavus Men-myco-93-63 as well as a preparation method and application thereof. In the invention, primers Ns8F and Ns8R are designed by utilizing an important negative regulator gene nsdB sequence in a Streptomyces coelicolor A3(2) antibiotic metabolic pathway, and genomic DNA (deoxyribonucleic acid) of Streptomyces roseoflavus Men-myco-93-63 is amplified to obtain a 1121bp segment; and two ends of a target sequence are extended by utilizing a primer anchoring polymerase chain reaction) (NPA-PCR ) method by a non-specific primer, thus obtaining the nsdBmgh gene of Streptomyces roseoflavus Men-myco-93-63. The preparation of the nsdBmgh gene provides an important basis for further researching and verifying the Streptomyces roseoflavus antibiotic metabolic pathway and regulatory mechanism, and establishes an important base for obtaining high-yield antibiotic gene engineering strains.

Description

technical field [0001] The present invention relates to the field of biotechnology, in particular to the production of Streptomyces roseoflavus (Streptomyces roseoflavus) Men-myco-93-63 nsdB mgh Genes, preparation methods and their applications. Background technique [0002] Cotton verticillium wilt is known as the "cancer" of cotton. As a typical soil-borne disease, its control has always been a worldwide problem. So far, there are neither high-resistant varieties nor ideal control methods. Cucumber powdery mildew is another important fungal plant disease in production. It is commonly known as white hair. It occurs all over the country. It is one of the main diseases of cucumbers in protected areas. In addition to harming cucumbers, it also harms zucchini, pumpkins, melons, wax gourds, Various cucurbit crops such as watermelon and loofah. Botrytis cinerea is a fungal disease caused by Botrytis, which mainly harms tomatoes, cucumbers, leeks, grapes, strawberries and othe...

Claims

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Application Information

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IPC IPC(8): C12N15/31C07K14/36C12N15/63C12N1/21C12N15/82A01N63/02A01P3/00C12N15/10C12R1/465C12R1/19
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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