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Promoter miR390 and applications thereof

A fragment and sequence technology, applied in the fields of application, angiosperms/flowering plants, DNA/RNA fragments, etc., can solve the problems of gene expression accumulation at the outer edge and affect plant growth and development, and achieve the effect of strong specific expression activity

Active Publication Date: 2012-06-27
INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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Problems solved by technology

However, because its expression does not distinguish between time and space, it will lead to excessive accumulation of outer edge gene expression, which will affect the normal growth and development of plants.

Method used

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  • Promoter miR390 and applications thereof
  • Promoter miR390 and applications thereof
  • Promoter miR390 and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1. Preparation of promoter and verification of specific expression

[0024] 1. Preparation and confirmation of the promoter

[0025] 1. Extraction of rice genomic DNA

[0026] (1) Take the leaves of Nipponbare rice, add liquid nitrogen, after fully grinding, pour the powder into a 1.5ml centrifuge tube;

[0027] (2) After the liquid nitrogen in the centrifuge tube is completely volatilized, immediately add 500ul DNA extraction buffer (0.1mol / LTris HCl (pH 8.0), 0.5mol / L NaCl, 0.05mol / L EDTA, 0.5% SDS), fully After mixing, incubate at 65°C for 30 minutes, during which the centrifuge tube is gently inverted up and down to fully mix the sample with the buffer;

[0028] (3) Centrifuge at room temperature for 15 minutes at 12,000 rpm;

[0029] (4) Transfer the supernatant to another centrifuge tube;

[0030] (5) Add an equal volume of tris-phenol and extract once;

[0031] (6) Extract again with equal volume of chloroform;

[0032] (7) Take the supernatant, add...

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Abstract

The invention discloses a promoter miR390 and applications thereof. The promoter is a deoxyribonucleic acid (DNA) fragment shown as in (1), (2) or (3) as follows: (1) a DNA fragment shown in a sequence 1 in a sequence table; (2) a DNA fragment which is hybridized with a DNA sequence limited by the DNA fragment shown in the sequence 1 in the sequence table under strict conditions and has same functions as the DNA sequence; and (3) a DNA fragment which has at least 90 percent of homology with the DNA sequence limited by the DNA fragment shown in the sequence 1 in the sequence table and has the same functions as the DNA sequence. Proved by experiments, the miR390 DNA fragment disclosed by the invention is a promoter, and moreover, the promoter has very strong specific expression activity, and is specifically expressed at the root cap of a plant. The promoter disclosed by the invention is capable of providing a facilitating device for function researches of specifically expressed genes and transgenic breeding of plants and has an important meaning.

Description

technical field [0001] The present invention relates to promoter miR390 and its application. Background technique [0002] Promoters can be divided into three categories according to their mode of action and function: constitutive promoters, inducible promoters, and tissue-specific promoters. Constitutive promoters can initiate expression in all tissues, with continuous expression and no time-space specificity, such as the CaMV35S promoter. Such promoters have been extensively studied and applied, and play an important role in genetic engineering. However, because its expression does not distinguish between time and space, it will lead to excessive accumulation of outer edge gene expression, which will affect the normal growth and development of plants. Therefore, searching for inducible or tissue-specific promoters plays an important role in gene function research and transgenic breeding. Contents of the invention [0003] One object of the present invention is to prov...

Claims

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Application Information

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IPC IPC(8): C12N15/113C12N15/11C12N15/63C12N5/10C12N1/15C12N1/19C12N1/21A01H5/00
Inventor 王秀杰王猛储成才曹守云
Owner INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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