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Flanking sequence of exogenous insert vector for transgenic rice and application of flanking sequence

A technology of transgenic rice and flanking sequences, applied in the field of biotechnology

Inactive Publication Date: 2012-08-08
SHANDONG AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, in the analysis of existing patents and documents, it is found that there are no articles or patent reports on the flanking sequence of the foreign insert of the transgenic rice line SK-2

Method used

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  • Flanking sequence of exogenous insert vector for transgenic rice and application of flanking sequence
  • Flanking sequence of exogenous insert vector for transgenic rice and application of flanking sequence
  • Flanking sequence of exogenous insert vector for transgenic rice and application of flanking sequence

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Cloning of the Side Sequence of the Foreign Insertion Vector of the Transgenic Rice Line SK-2

[0033] 1. Experimental materials

[0034] 1. Plant material

[0035] Transgenic rice: transgenic rice line SK-2.

[0036] 2. Experimental method and process

[0037] 1. Rice genomic DNA extraction and detection

[0038] 1) Extraction of rice genomic DNA

[0039] Rice leaves at the seedling stage were taken as materials for DNA extraction, and the total DNA of rice materials was extracted according to the operation manual of the column plant DNAout kit (Tianze Genomics Co., Ltd.).

[0040] 2) Genomic DNA detection of rice

[0041] Take 5ul of the extracted DNA solution, electrophoresis on 0.8% agarose gel, and judge the quality of the extracted DNA according to its brightness and band pattern. The concentration and purity of the extracted DNA were determined by UV spectrophotometer.

[0042] 2. Isolation of flanking sequences of transgenic rice line SK-2

[0043] 1) ...

Embodiment 2

[0063] Qualitative PCR detection method based on the flanking sequence of foreign insertion vector of transgenic rice line SK-2

[0064] 1. Experimental materials

[0065] 1. Plant material

[0066] Transgenic rice: transgenic rice line SK-2.

[0067] Conventional rice: Shengdao 301.

[0068] 2. Experimental method and process

[0069] 1. Plant genomic DNA extraction and detection

[0070] See "Plant Genomic DNA Extraction and Detection" in Example 1

[0071] 2. Strain-specific PCR detection based on the 3' and 5' flanking sequences

[0072] According to the side sequences of the 3' end and the 5' end determined in Example 1, primers were designed on the rice genome sequence part and the transformation vector sequence part respectively, as shown in Table 8. The results of PCR amplification showed that the combination of the 3' end and 5' end amplification primers of the transgenic rice line SK-2 obtained the expected amplified fragments of 383bp and 295bp, while the cont...

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Abstract

The invention relates to an exogenous insert vector flanking sequence for transgenic rice and application of the flanking sequence and belongs to the technical field of biology. According to the invention, a flanking sequence of an exogenous insert fragment of a transgenic rice variety SK-2 is cloned for the first time; the application that the flanking sequence of the exogenous insert fragment of the transgenic rice variety SK-2 can be used as a target DNA (Deoxyribose Nucleic Acid) amplification fragment is defined; and the variety specific qualitative PCR (Polymerase Chain Reaction) detection of a specific transgenic rice variety SK-2 is established. The cloned and separated flanking sequence of the exogenous insert fragment can be further used for establishing a transgenic rice variety SK-2 PCR detection method. The flanking sequence can be used for designing a specific primer for the amplification of target DNA and establishing the variety specific qualitative and quantitative PCR detection of the specific transgenic rice variety SK-2.

Description

technical field [0001] The present invention relates to a gene sequence in the field of biotechnology. Specifically, it relates to a flanking sequence of an exogenous insertion vector of a transgenic rice line SK-2. Background technique [0002] Since the first genetically modified tobacco crop came out in 1983, genetic engineering technology has developed rapidly, and new varieties of genetically modified crops have emerged. In recent years, genetically modified crops such as corn, soybean, rape, cotton, and tomato have been allowed to be planted and produced in many countries Widely used as raw material for food or feed processing. Because genetically modified products have a series of problems such as ecological risks and food safety, governments around the world and relevant international organizations have formulated corresponding safety management regulations, strengthened the standardized management of genetically improved crops, and implemented strict regulations o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12N15/10C12Q1/68
Inventor 杨正友李文华赵凤春田园
Owner SHANDONG AGRICULTURAL UNIVERSITY
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