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Specific expression promoter for plant embryo and application thereof

A specificity and promoter technology, applied in application, plant products, botanical equipment and methods, etc., to achieve the effect of improving expression and accumulation level, increasing added value of science and technology, and having great application prospects

Inactive Publication Date: 2011-11-23
INST OF BOTANY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The product encoded by the rice Os11S (GeneID: 4329281) gene is a seed storage protein, and there is no research report on the tissue-specific expression of the Os11S gene in seeds

Method used

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  • Specific expression promoter for plant embryo and application thereof
  • Specific expression promoter for plant embryo and application thereof
  • Specific expression promoter for plant embryo and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Embodiment 1, the acquisition of plant embryo-specific expression promoter (Os11S promoter)

[0038] The cDNA sequence of an 11S protein gene (gene name Os02g0456100, Os11S) was queried from the rice database, the upstream sequence of the Os11S gene was searched from GenBank, and primers were designed to amplify the Os11S promoter. To facilitate vector construction, two restriction sites (underlined) were sequentially added to the primers. Os11SHindF: 5'-AA AAGCTT GAAGTAGGCTGTGTTGCAGAGG-3'( Hind III ) is the forward primer of Os11S promoter, Os11SSalR: 5'-GC GTC GAC AATTCTAGTCACTGCAATAAGTAG-3'( Sal I ) is the reverse primer.

[0039] The CTAB method was used to extract a small amount of genome from the leaves of rice (wild type rice Taichung No. 65, which was bred in Taiwan Province of my country in 1936; Qu Leqing et al., Acta Botany, 2001, 43: 1167-1171; preserved by the Institute of Botany, Chinese Academy of Sciences) Using DNA as a template and Os11SHindF a...

Embodiment 2

[0040] Embodiment 2, plant embryo-specific expression promoter (Os11S promoter) expression vector construction and genetic transformation

[0041] 1. Construction of plant expression vector with Os11S promoter fused to GUS gene

[0042] pGPTV-35S-HPT was constructed with reference to the method described in the literature (Qu and Takaiwa, Plant Biotech J 2004, 2: 113-125), and was preserved by the Institute of Botany, Chinese Academy of Sciences. The pMD-Os11S plasmid and the binary expression vector pGPTV-35S-HPT were digested with Sal I and Hind III. A 2230bp enzyme-digested fragment containing the Os11S promoter was recovered, and the fragment was ligated between the Sal I and Hind III enzyme recognition sites of pGPTV-35S-HPT. On the basis of PCR identification, the obtained recombinant plasmid was identified by double enzyme digestion with Sal I and Hind III, and a fragment containing 2.2 kb Os11S promoter was obtained. The double-enzyme digestion identification showed ...

Embodiment 3

[0047] Example 3, Functional Verification of Plant Embryo-Specific Expression Promoter (Os11S Promoter)

[0048] Transgenic Os11S-pGPTV-35S-HPT rice T 0 Plants were subjected to histochemical staining. Specific steps: transfer Os11S-pGPTV-35S-HPT rice T 0 Part of the leaves, roots, stem tissues of the generation plants, and the seeds at the filling stage 11 days, 15 days, and 17 days after flowering that were cut longitudinally from the middle with a scalpel were soaked in GUS reaction solution (0.1M NaPO 4 Buffer, pH 7.0, 10 mM EDTA, pH 7.0, 5 mM potassium ferricyanide, 5 mM potassium ferrocyanide, 1.0 mMX-Gluc, 0.1% Triton X-100), react at 37°C. The stained tissues were preserved in 70% ethanol, observed, and photographed under a dissecting microscope. The results showed that GUS expression was not observed in the roots, stems and leaves of the transgenic Os11S-pGPTV-35S-HPT rice. Image 6 Shown; Embryos of seeds 11 days after flowering turned blue; seeds of 15 days afte...

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Abstract

The invention discloses a specific expression promoter for a plant embryo and application thereof. The promoter is derived from (1) a DNA (Deoxyribonucleic Acid) sequence shown as a sequence 1 in a sequence table, (2) a DNA sequence having over 70 percent of homology with the DNA sequence defined by the sequence 1 in the sequence table and having the function of a specific expression promoter forthe plant embryo, and (3) a nucleotide sequence which can be hybridized with the DNA sequence defined by the sequence 1 in the sequence table under a highly serious condition. The promoter can be used for promoting specific expression of an exogenous gene in the plant embryo, and is suitable for any seed plant.

Description

technical field [0001] The invention relates to a plant embryo-specific expression promoter and its application. Background technique [0002] The growth, development and physiological process of organisms are the result of orderly and specific expression of many genes under the action of external environmental factors. Gene expression is finely regulated by various physical and chemical factors in the organism at the DNA level, transcription level, post-transcription level, translation level, post-translation processing, transportation and positioning. This process involves promoters, terminators, UTR The role of a series of regulatory elements such as sequences. In recent years, molecular biology studies have shown that the regulation of transcription level is the initial step in the regulation of gene expression, and it is also one of the important ways of gene expression regulation. Promoter refers to a specific DNA sequence that RNA polymerase and some trans-acting fa...

Claims

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Application Information

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IPC IPC(8): C12N15/113C12N15/63C12N5/10C12N1/15C12N1/19C12N1/21A01H5/00
Inventor 曲乐庆徐秀萍董祥柏柴志坚
Owner INST OF BOTANY CHINESE ACAD OF SCI
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