Primer groups, kit and method for detecting specific sequence of glyphosate-resistant transgenic soybean transformant

A technology of genetically modified soybeans and detection kits, which is applied in the directions of biochemical equipment and methods, recombinant DNA technology, microbial measurement/inspection, etc. Sexual distinction and other issues, to achieve the effect of easy large-scale promotion and application, visualization of results, and avoiding false positives and false negatives

Inactive Publication Date: 2012-11-07
GUANGDONG TESTING INST OF PROD QUALITY SUPERVISION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, since the cp4-epsps gene, as an excellent resistance gene for the benefit of human beings, has been transferred into multiple crops or different lines of the same crop to form different lines of transgenic crops, the detection method obviously cannot effectively and specifically distinguish the genetically modified crops with Genetically modified crops and their lines with the same resistance to glyphosate
In 2011, Cao Yicheng and others also established a LAMP detection kit for detecting transgenic soybeans. This kit is for the amplification detection of exogenous EPSPS-NOS gene, but the EPSPS-NOS gene sequence does not only exist in glyphosate-resistant Glyphosate-resistant GM soybean components cannot be specifically detected in GM soybeans

Method used

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  • Primer groups, kit and method for detecting specific sequence of glyphosate-resistant transgenic soybean transformant
  • Primer groups, kit and method for detecting specific sequence of glyphosate-resistant transgenic soybean transformant
  • Primer groups, kit and method for detecting specific sequence of glyphosate-resistant transgenic soybean transformant

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1: Development of glyphosate-resistant transgenic soybean detection kit

[0043] The glyphosate-resistant transgenic soybean detection kit of the present invention consists of (1) Bst DNA polymerase; (2) primer set 1; (3) primer set 2; (4) reaction solution; (5) dye (SYBR Green) (6) Internal control; (7) Positive control composition, in which Bst DNA polymerase, reaction solution, color development solution, etc. can be purchased from the corresponding company, and the detection primer set, internal control and positive control are self-developed.

[0044] 1. Experimental materials

[0045] glyphosate resistant genetically modified soybeans

[0046] 2. Reagents

[0047] Bst DNA polymerase, 10× Bst buffer was purchased from New England Biolabs, USA; betaine was purchased from Sigma, USA; dNTP, MgSO4 analytically pure, Premix Taq DNA polymerase, pMD18-T vector, DNA molecular weight marker, and transgenic genomic DNA extraction kit (GMO Detection Ver.2.0...

Embodiment 2

[0080] Example 2: Application of glyphosate-resistant transgenic soybean detection kit

[0081] 1. Experimental materials

[0082] Glyphosate-resistant genetically modified soybeans.

[0083] Non-GMO soybeans.

[0084] Other genetically modified materials: genetically modified rice TT51-1, genetically modified rapeseed RT73, genetically modified sugar beet H7-1, genetically modified corn MON810, and genetically modified corn Bt176.

[0085] 2. Reagents

[0086] Bst DNA polymerase, 10× Bst Buffer was purchased from New England Biolabs, USA; betaine was purchased from Sigma, USA; analytically pure dNTP, MgSO4, and transgenic genomic DNA extraction kit (GMO Detection Ver.2.0) were purchased from Treasure Bioengineering (Dalian) Co., Ltd.; Synthesized by Weijieji Bioengineering Technology Service Co., Ltd.

[0087] 3. Main instruments and equipment

[0088] Gel-type gel imaging system (Bio-Rad Laboratories, Inc.)

[0089] Biophotometer plus UV spectrophotometer (Eppen...

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Abstract

The invention discloses primer groups, a kit and a method for detecting a specific sequence of a glyphosate-resistant transgenic soybean transformant. Two groups of LAMP (loop-mediated isothermal amplification) primers are designed, the primer group 1 is used for detecting an internal reference gene Lectin and can detect soybean ingredients in a sample and simultaneously exclude false negative results, and the primer group 2 is used for detecting the specific sequence of the glyphosate-resistant transgenic soybean transformant, has high specificity and can effectively detect glyphosate-resistant transgenic soybean ingredients, wherein the sequence is a boundary sequence which connects an inserted exogenous gene with a plant gene. When the kit is used for the detection and analysis of a sample, a positive control, a blank control and an internal control are respectively set, so that the production of false positives and false negatives can be avoided. Special equipment is not required for detection, and the detection cost is low; and the detection results can be judged by visual observation of changes in color of a reaction solution, the identification is simple and convenient to perform, the results are visualized, and the primer groups, the kit and the method disclosed by the invention are suitable for field fast detection and grassroots screening of a large number of samples and easy to realize large-range popularization and application.

Description

technical field [0001] The invention belongs to the field of biological detection, and in particular relates to a primer set, a kit and a method for detecting the specific sequence of a glyphosate-resistant transgenic soybean transformant. Background technique [0002] With the rapid development of genetically modified organisms, their safety has aroused intense debate and widespread concern around the world. In order to protect consumers' right to know and right to choose, many countries and regions in the world have enacted corresponding laws and regulations to strengthen the management of import and export of genetically modified products, which brings new challenges to the genetically modified testing work that provides technical support for the safety supervision of genetically modified organisms. Come to greater challenges. There are many methods for detecting genetically modified products, which are mainly divided into two categories: detection of exogenous proteins ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
Inventor 刘唐书肖维威周琳华佘之韵梁宇斌张宝
Owner GUANGDONG TESTING INST OF PROD QUALITY SUPERVISION
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