Rice-derived endosperm specific expression promoter and application thereof

An expression cassette and molecular technology, applied in the application, introduction of foreign genetic material and modified cells, angiosperms/flowering plants, etc., can solve the problem of consumption of bioenergy, adverse effects of biological growth and development on metabolites, and expression intensity that cannot meet the requirements of the transgenic industry To meet the needs of globalization and other issues, to achieve great application prospects, increase the added value of science and technology, and improve the effect of seed quality

Active Publication Date: 2014-03-26
INST OF BOTANY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Although the widely used promoters (including CaMV35S, ubiquitin1, and Actin1) have high expression efficiency, they can express foreign genes in almost all tissues because their expression is not limited by time and space. During the expression process, it will drive the expression of genes in other tissues outside the seed, which will not only consume bioenergy, but also lead to the synthesis of some metabolites that may adversely affect the growth and development of organisms
Moreover, the expression intensity of the above-mentioned promoters cannot meet the needs of transgenic industrialization.

Method used

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  • Rice-derived endosperm specific expression promoter and application thereof
  • Rice-derived endosperm specific expression promoter and application thereof
  • Rice-derived endosperm specific expression promoter and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1, the acquisition of plant seed endosperm-specific expression promoter SSP-1

[0045] Genomic DNA was extracted from rice Kitaake leaves by CTAB method, and the promoter SSP-1 was amplified by PCR using it as a template and P1 and P2 as primers. The reaction system is: 1 μl of 10 μM forward and reverse primers, 10×Ex Buffer 2 μl, genomic DNA 1 μl (10 ng), ExTaq (TaKaRa) 0.5 unit, add ultrapure water to 20 μl; the reaction program is: 94 ° C for 5 minutes, then 94 ° C 1min, 55°C 1min30sec, 72°C 2min30sec, 30 cycles, finally 72°C, 10min. The target band of 2.4kb was amplified by PCR. The amplified product was recovered, directly connected to the pMD18-T vector (purchased from TaKaRa Company), and sequenced. The results showed that the promoter SSP-1 (2416bp) shown in SEQ ID No.1 was amplified. The recombinant vector containing the promoter SSP-1 was named as pMD-SSP-1 by sequencing detection.

[0046] Wherein, the forward primer (P1) sequence of PCR is: 5'-GG ...

Embodiment 2

[0047] Example 2, Plant Seed Endosperm-Specific Expression Promoter SSP-1 Promotes the Specific Expression of Exogenous Genes in Rice Seed Endosperm

[0048] 1. Construction of a plant expression vector containing an exogenous gene expression cassette promoted by SSP-1

[0049] Digest pMD-SSP-1 with Sal I and Hind III, recover a 2416bp enzyme-cut fragment containing the promoter SSP-1, and insert the fragment between the Sal I and Hind III enzyme recognition sites of pGPTV-35S-HPT During the period, the recombinant expression vector obtained by replacing the fragment between the Sal I and Hind III enzyme recognition sites of pGPTV-35S-HPT with the promoter SSP-1 shown in SEQ ID No.1 was named pGPTV-SSP-1- Gus. The schematic diagram of the structure of pGPTV-SSP-1-GUS is shown in figure 1 As shown, it contains the GUS gene expression cassette, which is composed of the promoter SSP-1, the exogenous gene GUS gene (UidA) driven by the promoter SSP-1, and the terminator Tnos. Sa...

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Abstract

The invention discloses a rice-derived endosperm specific expression promoter and application thereof. The promoter is a DNA molecule as shown in the following a), b) or c): a) the DNA molecule with the nucleotide sequence of SEQ ID No.1; b) the DNA molecule with 70% or more than 70% of identity with the nucleotide sequence defined by a) and with a promoter function; and c) the DNA molecule which is hybridized with the nucleotide sequence defined by a) or b) under the highly strict condition and has a promoter function. By adoption of the promoter provided by the invention, the expression and accumulation level of an exogenous gene to plant seed endosperm can be increased; the seed quality can be improved; protein or oligopeptide with physiological activity can be led into the seed to create a health-care type functional novel variety; and the seed can serve as a bioreactor to produce useful foreign protein or edible vaccine so as to increase the technological additional value of agricultural products.

Description

technical field [0001] The invention relates to an endosperm-specific expression promoter derived from rice and its application. Background technique [0002] The latest development of plant biotechnology not only realizes the improvement of traditional agronomic shapes (such as increasing crop yield, enhancing disease resistance, insect resistance, herbicide resistance, improving quality, etc.), but also makes plants become bioreactors for biomedicine and industrial products (Daniell et al., Trends Plant Sci.2001,6:219-226; Giddings et al., Nature Biotech.2000,18:1151-1156; Hood and Jilka, Curr.Opin.Biotechnol.1999,10:382- 386; Hood and Woodard, Plants as Factories for Protein Production, 2002, pp.119-135. Netherlands: Kluwer Academic). For most gramineous crops, due to its high yield, low production cost, storage resistance and easy control of production scale, it is an ideal carrier for the second generation of transgenic products. In recent years, the use of rice seeds...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12N15/82C12N5/10C12N1/15C12N1/19C12N1/21A01H5/10
Inventor 曲乐庆董祥柏
Owner INST OF BOTANY CHINESE ACAD OF SCI
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