Specific expression promoter of rice seed aleurone layer and application thereof

A technology in the promoter and sequence listing, which is applied in the direction of using vectors to introduce foreign genetic material, DNA/RNA fragments, recombinant DNA technology, etc., and can solve problems such as the inability to meet the needs of genetic engineering to improve the quality of rice

Active Publication Date: 2013-07-10
INST OF BOTANY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These known promoters are far from meeting the needs of genetic engineering to improve rice quality, so the present invention has important application prospects

Method used

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  • Specific expression promoter of rice seed aleurone layer and application thereof
  • Specific expression promoter of rice seed aleurone layer and application thereof
  • Specific expression promoter of rice seed aleurone layer and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Embodiment 1, the acquisition of promoter

[0029] 1. Screening of rice seed aleurone layer-specific expression genes

[0030] 1. Fluorescent quantitative PCR screening of transcription factors specifically expressed in the rice seed aleurone layer

[0031] Select rice leaves, spikelets, roots, rice seeds of different developmental stages (4, 7, 11, 14, 21 days) and hand-separated rice seed starch endosperm and aleurone layer (11 days of seeds), respectively extract mRNA, cDNA was synthesized by reverse transcription, and the expression profile of rice NF-YB1 gene (gene number: LOC_Os02g49410) in different tissues and developmental stages was identified by fluorescent quantitative PCR, and the specific expression of NF-YB1 gene in rice aleurone layer was preliminarily determined ( figure 1 ).

[0032] 2. Tissue in situ hybridization to verify the specific expression of NF-YB1 gene in the rice seed aleurone layer

[0033] The seeds of different development stages of r...

Embodiment 2

[0046] Example 2, functional verification of the promoter

[0047] 1. Obtaining of transgenic P-NF-YB1 rice

[0048] 1. Obtaining the recombinant vector

[0049] The 3018bp PCR product P-NF-YB1 obtained by Example 1 and the expression vector pPLV15 (Gene bank No. JF909468, the literature that records the pPLV15 are as follows: A Versatile Set of Ligation-Independent Cloning Vectors for Functional Studies in Plants. Author: Bert De Rybel, Willy van den Berg, Annemarie S. Lokerse, Che-Yang Liao, Hilda van Mourik, Barbara Mo¨ller, Cristina I. Llavata-Peris, and Dolf Weijers, Plant physiology, (2011), 156:1292-1299 ; The public can obtain it from the Institute of Botany, Chinese Academy of Sciences, and the structural diagram of the carrier is as follows image 3 (shown) is a method that relies on the cohesive ends of more than 20 bases for complementary pairing: first cut the vector pPLV15 with HpaI, and then use T 4 DNA polymerase digests the PCR product and the vector pPLV15...

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Abstract

The invention discloses a specific expression promoter of a rice seed aleurone layer and application of the rice seed aleurone layer, and provides a deoxyribonucleic acid (DNA) segment which is a DNA molecule shown in 1) or 2) or 3), wherein 1) is the DNA molecule shown in a sequence 1 of a sequence table; 2) is the DNA molecule hybridized with a DNA sequence defined by 1) under a strict condition and having a promoter function; and 3) is the DNA molecule having homology of more than 70 percent with the DNA sequence defined by 1) and having a promoter function. The experiment shows that the promoter P-NF-YB1 is found, rice is introduced, and the expression of glucuronidase (GUS) gene is driven in the rice. The result shows that the rice seed aleurone layer specially expresses that the promoter is a tissue specificity promoter; and the promoter has wide application and market prospect in the field of agriculture.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a rice seed aleurone-specific expression promoter and application thereof. Background technique [0002] Promoter refers to a DNA sequence in a gene that can accurately and effectively initiate transcription, usually located upstream of the gene. The promoter of higher plants is generally composed of two parts: one part is required to form a general transcription structure, usually called the core promoter region, including the transcription initiation point and the adjacent TATA box; the other part is the region that determines the specificity and activity of gene transcription , consisting of multiple conserved sequences, the position, type and copy number of these conserved sequences in different promoters are quite different. Promoters are involved in regulating the expression of target genes in specific tissues, developmental stages and environmental conditions. Therefore, und...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12N15/63C12N5/10C12N1/15C12N1/19C12N1/21C12N15/11
Inventor 刘春明白爱宁张晋丹
Owner INST OF BOTANY CHINESE ACAD OF SCI
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