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Promoter AFB4 and application thereof

A technology in fragments and sequence lists, applied in applications, recombinant DNA technology, angiosperms/flowering plants, 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

Inactive Publication Date: 2012-05-23
INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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  • Abstract
  • Description
  • Claims
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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 AFB4 and application thereof
  • Promoter AFB4 and application thereof
  • Promoter AFB4 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1. Preparation of promoter and verification of specific expression

[0026] 1. Preparation and confirmation of promoter

[0027] 1. Extraction of Arabidopsis genomic DNA

[0028] (1) Take Arabidopsis thaliana leaves, add liquid nitrogen, and after fully grinding, pour the powder into a 1.5ml centrifuge tube;

[0029] (2) After the liquid nitrogen in the centrifuge tube has completely evaporated, 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 was gently inverted up and down to mix the sample and buffer well;

[0030] (3) Centrifuge at 12,000 rpm for 15 minutes at room temperature;

[0031] (4) transfer the supernatant to another centrifuge tube;

[0032] (5) adding an equal volume of tris-phenol and extracting once;

[0033] (6) extract again with equal volume of chloroform;

[0034] (7) Take the supernatan...

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Abstract

The invention discloses a promoter AFB4 and application thereof. The promoter is a DNA segment shown in the following 1), 2), or 3): 1) a DNA segment shown in a sequence 1 of a sequence table; 2) a DNA segment which is hybridized with the DNA sequence defined in 1) under a strict condition and has the same function as the DNA sequence defined in 1); and 3) a DNA segment which has over 90 percent of homology and the same function as the DNA sequence defined in 1). Experiments prove that the AFB4 DNA segment is a promoter which has very high specific expression activity and specifically expresses on a vascular cylinder of a main root of a plant, combined with a branch root, and the jagged edges of leaves. The promoter provides a beneficial tool for a gene functional research on specific expression and the transgenic breeding of the plant and has great significance.

Description

technical field [0001] The present invention relates to the promoter AFB4 and its use. 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, have continuous expression, and have no spatiotemporal 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 is not divided in time and space, it will lead to the excessive accumulation of outer edge gene expression, which will affect the normal growth and development of plants. Therefore, finding inducible or tissue-specific promoters plays an important role in gene function research and transgenic breeding. SUMMARY OF THE INVENTION [0003] An object of the present invention is to provide a DNA fr...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/113C12N15/11C12N15/63C12N1/15C12N1/19C12N1/21C12N5/10A01H5/00
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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