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Agrobacterium tumefaciens-mediated genetic transformation method of Penicillium italicum

A genetic transformation method, the technology of Agrobacterium tumefaciens, applied in the direction of using vectors to introduce foreign genetic material, recombinant DNA technology, etc., can solve the problems of fruit rot and economic loss in agricultural production, and achieve high transformation rate, simple operation and stable transformation Effect

Inactive Publication Date: 2017-09-19
HUAZHONG AGRI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Penicillium italicum causes a large number of fruit rots during the postharvest storage and transportation of citrus, which brings huge economic losses to agricultural production

Method used

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  • Agrobacterium tumefaciens-mediated genetic transformation method of Penicillium italicum
  • Agrobacterium tumefaciens-mediated genetic transformation method of Penicillium italicum
  • Agrobacterium tumefaciens-mediated genetic transformation method of Penicillium italicum

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Embodiment Construction

[0033] The present invention will be described in detail below by way of examples.

[0034] 1. Experimental materials

[0035] 1.1 Strains and vectors

[0036] Agrobacterium tumefaciens GV3101, Agrobacterium tumefaciens AGL-1, Escherichia coli DH5α

[0037] The pMD19-T vector, plasmid pSKH, and plasmid pCAMBIA3300 were used to construct the knockout vector pacC3300

[0038] 1.2 Medium

[0039] Potato Dextrose Agar Medium (PDA): Potato 200g, take the filtrate after boiling, add 10g of sucrose, 20g of agar, for the cultivation of Penicillium italicum.

[0040] Beef extract peptone liquid medium (LB): Tryptone 10g, yeast extract powder 5g, NaCl 10g, adjust pH to 7.4 with NaOH, add ddH2O to 1L.

[0041] Beef peptone solid medium (LBA): add 20g agar on the basis of LB medium

[0042] Table 1 Medium composition list

[0043]

[0044] Note: Glucose, AS and MES need to be sterilized by filtration.

[0045] 2. Construction of knockout vector

[0046] 2.1 Construction of the ...

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Abstract

The invention discloses an Agrobacterium tumefaciens-mediated genetic transformation method of Penicillium italicum and belongs to the field of genetic transformation. The method of the invention is simple to perform, high in transformation rate and good in transformation stability, provides efficient genetic transformation of Penicillium italicum successfully, makes it possible to perform homologous knockout, overexpression and heterologous expression in Penicillium italicum, and lays basis for the genetic improvement of this strain by genetic engineering means and the study on pathogenesis of citrus infection by the strain.

Description

technical field [0001] The invention relates to a genetic transformation method of penicillium italicum, belonging to the field of genetic transformation. Background technique [0002] Citrus fruits are very popular in the daily sales of fruits for their unique aroma and taste and play an important role in nutrition supply and health promotion. Penicillium italicum causes a large number of fruit rots during postharvest storage and transportation of citrus, which brings huge economic losses to agricultural production. Through Agrobacterium-mediated genetic transformation technology, the function of specific genes of Penicillium italicum can be explored, so as to further explore the pathogenic mechanism of Penicillium italicum and control the disease more effectively. [0003] Agrobacterium tumefaciens-mediated transformation is currently the most widely used genetic transformation method. Agrobacterium tumefaciens is a special plant pathogen that can be transferred to the h...

Claims

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Application Information

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IPC IPC(8): C12N15/80
Inventor 彭丽桃李倩如程晨杨书珍
Owner HUAZHONG AGRI UNIV
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