Exogenous insertion element flanking sequences of transgenic rice strain 223F-S21 and application thereof

A technology of 223F-S21 and transgenic rice, which is applied in the direction of DNA/RNA fragments, microbe determination/inspection, DNA preparation, etc., and can solve the problem of side sequence of foreign insert fragment without transgenic rice line 223F-S21, etc.

Inactive Publication Date: 2014-05-07
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the analysis of existing patents and documents found that there are no articles or patent reports on the flanking sequence of the foreign insert of the transgenic rice line 223F-S21

Method used

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  • Exogenous insertion element flanking sequences of transgenic rice strain 223F-S21 and application thereof
  • Exogenous insertion element flanking sequences of transgenic rice strain 223F-S21 and application thereof
  • Exogenous insertion element flanking sequences of transgenic rice strain 223F-S21 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Example 1, Cloning of the flanking sequence of the exogenous insertion vector of the transgenic rice line 223F-S21

[0059] 1. Experimental materials

[0060] 1. Plant material

[0061] Transgenic rice: transgenic rice line 223F-S21.

[0062] 2. Enzymes and reagents

[0063] Molecular biology reagents, such as Taq enzyme, TaKaRaLA Taq, pMD18-T, and DNA Marker were purchased from Dalian Bao Biological Engineering Co., Ltd.

[0064] Other biochemical reagents are imported or domestic analytically pure. Primers and degenerate primers were synthesized by Shanghai Sangong.

[0065] 3. Experimental equipment

[0066] PCR amplification (Eppendorf co.), nucleic acid electrophoresis instrument (DYY-6C, Beijing Liuyi Instrument Factory)

[0067] Other instruments include: centrifuge, electric heating constant temperature water tank, electronic balance, incubator, etc.

[0068] 2. Experimental method and process

[0069] 1. Rice genomic DNA extraction and detection

[007...

Embodiment 2

[0109] Qualitative PCR detection of the flanking sequence of the foreign insert in transgenic rice 223F-S21

[0110] 1. Experimental materials

[0111] 1. Plant material

[0112] Transgenic rice: Transgenic rice line 223F-S21

[0113] Conventional rice: Xiushui 134

[0114] 2. Enzymes and reagents

[0115] Molecular biology reagents, such as Taq enzyme, TaKaRaLA Taq, pMD18-T, and DNA Marker were purchased from Dalian Bao Biological Engineering Co., Ltd.

[0116] Other biochemical reagents are imported or domestic analytically pure. Primers and degenerate primers were synthesized by Shanghai Sangong.

[0117] 3. Experimental equipment

[0118] PCR amplification (Eppendorf co.), nucleic acid electrophoresis instrument (DYY-6C, Beijing Liuyi Instrument Factory)

[0119] Other instruments include: centrifuge, electric heating constant temperature water tank, electronic balance, incubator, etc.

[0120] 2. Experimental method and process

[0121] 1. Rice genomic DNA extrac...

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PUM

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Abstract

The invention discloses an exogenous insertion element flanking sequence of a transgenic rice strain 223F-S21, wherein the flanking sequence is a 3-end flanking sequence and is shown as SEQ ID NO. 1. The invention further discloses an exogenous insertion element flanking sequence of another transgenic rice strain 223F-S21, wherein the flanking sequence is a 5-end flanking sequence and is shown as SEQ ID NO. 2. According to the invention, the exogenous insertion element flanking sequences of the transgenic rice strain 223F-S21 are cloned for the first time, and the situation that the exogenous insertion element flanking sequences of the transgenic rice strain 223F-S21 can be used as targeted DNA amplified fragments to establish sensitive and specific the strain specific qualitative PCR of the transgenic rice strain 223F-S21 is definite. By adopting the exogenous insertion element flanking sequences cloned and separated by the invention, a transgenic-strain specific PCR detection method can be further established.

Description

technical field [0001] The present invention relates to the gene sequence in the field of biotechnology, in particular to the side sequence of an exogenous insert segment of a transgenic rice line 223F-S21. Background technique [0002] With the continuous development of genetic engineering technology, new varieties of genetically modified crops such as corn, soybean, rapeseed, cotton, tomato, etc. continue to emerge, and many countries have allowed the planting and production of genetically modified crops. Rice is one of the most important food crops in my country. With the in-depth research and wide application of transgenic technology in rice, several transgenic rice strains have been released to the environment and applied for commercial planting. Since genetically modified products have a series of problems such as ecological risks and food safety, countries around the world have formulated corresponding safety management laws and regulations. Strain-specific detection...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12N15/10C12Q1/68
Inventor 杨永义沈志成
Owner ZHEJIANG UNIV
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