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Gibberella fujikuroi electroporation genetic transformation method

A genetic transformation method, the technology of Gibberella Fujikura, applied in the field of molecular biology research of Gibberella, can solve the problems of many transformation steps and low efficiency, and achieve the effect of improving the synthesis ability

Inactive Publication Date: 2007-07-11
ZHEJIANG UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The spore transformation method is limited to Gibberella strains that can produce spores. The PEG-mediated protoplast transformation method is widely used, but the transformation steps are many and the efficiency is not high.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1: Genetic transformation of exogenous hygromycin resistance gene to Gibberella fujikuroi by electroporation

[0032] Using the method described in the Summary of the Invention, the exogenous plasmid pAN7-1 containing the hygromycin resistance gene was introduced into Gibberella, and the stably transformed engineered bacteria were obtained through hygromycin screening, and the transformation efficiency was higher than that mediated by lithium acetate. The spore transformation method is simpler than the transformation method mediated by polyethylene glycol PEG. The specific method is as follows:

[0033] 1. Preparation of Gibberella protoplasts:

[0034] 1) Activating Gibberella fujikura species on a PDA medium plate at a culture temperature of 26-28°C.

[0035] The final concentration of PDA medium is: potato, 200g / L; sucrose, 20g / L; agar powder, 15g / L; pH is natural. Sterilize at 121°C for 18 minutes.

[0036]2) Take a small piece and inoculate it into MYG ...

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Abstract

The invention discloses a genetic conversing method of exogenous gene for Gibberalla fujikuroi through electroporation technique, which is characterized by the following: utilizing cell wall degraded enzyme of fungi to dispose the cell wall; obtaining the protoplast to purify; blending purified protoplast and fitful exogenous gene / DNA solution; proceeding electric shock effect; making protoplasm adsorb the exogenous DNA.

Description

(1) Technical field [0001] The invention relates to the electroporation genetic transformation technology of Gibberella fujikuroi, which is applicable to the fields of genetic engineering of Gibberella fujikuroi, molecular biology research of Gibberella and the like. (2) Background technology [0002] Gibberella fujikuroi is a gibberellin-producing fungus that causes bakanae disease due to excessive production of gibberellin in rice. The disease occurs in various rice-producing areas in my country and poses a serious threat to rice production. Gibberellin is a plant growth regulator, which plays a very important role in plant growth and development, and is widely used in fruits, vegetables, rice and other crops. Gibberellin can also be used in beer production industry, clinical medicine and so on. [0003] Gibberella has become one of the filamentous fungi with good development prospects. At present, many studies are trying to improve strains through genetic engineering. A...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/87C12N1/15C12N15/80
Inventor 朱廷恒王渭霞汪琨崔志峰裘娟萍
Owner ZHEJIANG UNIV OF TECH
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