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Corn callus specific promoter and cloning method and application thereof

A callus and promoter technology, applied in recombinant DNA technology, introduction of foreign genetic material, DNA/RNA fragments using vectors, etc. Avoid unnecessary wasteful effects

Inactive Publication Date: 2011-06-15
ANHUI AGRICULTURAL UNIVERSITY
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Problems solved by technology

However, the above-mentioned elimination of the screening marker gene at the genetic level has the disadvantages of being cumbersome and complicated, and the transformation cycle is long.

Method used

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  • Corn callus specific promoter and cloning method and application thereof
  • Corn callus specific promoter and cloning method and application thereof
  • Corn callus specific promoter and cloning method and application thereof

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Embodiment Construction

[0011] The present invention will be further described below in conjunction with specific embodiments.

[0012] The methods used in the following examples are conventional methods unless otherwise specified. The primers used were all synthesized by Shanghai Jierui Bioengineering Co., Ltd., and the sequencing was performed by Invitrogen Company. Dicer enzymes were purchased from Zibao Bioengineering Co., Ltd., plasmid extraction kits and gel recovery kits were purchased from Quanshijin Biotechnology Co., Ltd., and the methods were carried out according to the instructions of the kits.

[0013] 1. Genes specifically expressed in maize callus wrky the acquisition

[0014] First, search for the gene with the keyword "wrky" on the NCBI website (http: / / www.ncbi.nlm.nih.gov), and obtain the protein sequence of the Arabidopsis wrky gene. A local corn protein database was established, and the protein sequence of the obtained Arabidopsis wrky gene was compared by BlastP (E-value=0.00...

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Abstract

The invention discloses a corn callus specific promoter and a cloning method and application thereof. A WRKY gene is obtained by utilizing bioinformatics analysis, the sequence comparison of the gene and the genome is performed, a primer is designed to perform polymerase chain reaction (PCR) and obtain the promoter region of the gene 5', the promoter is 2880bp and has the sequence shown in the (400)1 item of the sequence table; the analysis of the promoter shows that the promoter contains a plurality of acting elements; and the promoter is subcloned to the vector with GUS gene, expression of the GUS gene is started, and the vector is transformed in rice. The staining examine of each tissue of rice shows that the specificity of the promoter can be expressed in callus, thus the promoter hasimportant application value in genetic engineering and the gene function researches.

Description

technical field [0001] The invention relates to a plant promoter, its cloning method and application thereof, in particular to a maize callus-specific promoter, its cloning method and its application in cultivating safe transgenic plants. Background technique [0002] Screening marker genes play an important role in plant genetic engineering. Transformed cells can be screened out from a large number of untransformed cells during the plant transgenic process by using the screenable marker gene. However, the selection of genes in transgenic plants has caused people to worry about a series of biosafety aspects, such as whether the selection marker gene encodes toxic substances and allergens, whether it is not conducive to changes in plant metabolism, and whether it will reduce the efficacy of therapeutic drugs , and whether it will migrate between similar species and pathogens. How to remove screening marker genes from transgenic plants has become a concern of people. At pres...

Claims

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Application Information

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IPC IPC(8): C12N15/113C12N15/82
Inventor 朱苏文程备久项艳王莹唐秀丽
Owner ANHUI AGRICULTURAL UNIVERSITY
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