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Genetically-modified crop detection method

A detection method and genetically modified technology, applied in the determination/inspection of microorganisms, biochemical equipment and methods, recombinant DNA technology, etc., can solve problems such as labor-intensive and impractical

Pending Publication Date: 2018-08-31
NISSHIN SEIFUN GRP INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, such a method is labor-intensive and impractical

Method used

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  • Genetically-modified crop detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] [Example 1: Detection of soybean-derived and cotton-derived DREB genes by PCR and agarose gel electrophoresis]

[0063] (method)

[0064] Using the DNA extracted from each of the following three test samples as templates, PCR was performed using the following primer pairs. Thereafter, the presence or absence of amplification of each sample was examined by agarose gel electrophoresis.

[0065] (sample)

[0066] Wheat: Wheat produced in Japan

[0067] Soybeans: Dried seeds of commercially available Japanese soybeans

[0068] Cotton: dried seeds of commercially available cotton

[0069] After pulverizing each sample with a Multi Beads Shocker (Yasui Instrument Co., Ltd.), DNA was extracted using a DNeasy Plant Maxi kit (manufactured by QIAGEN).

[0070] (primer pair)

[0071] In order to design primers used in PCR, a gene sequence database (National Center for Biotechnology Information) (http: / / www.ncbi.nlm.nih.gov / ) was used.

[0072] As the DNA sequence of the soybe...

Embodiment 2

[0085] [Example 2: Detection of DREB genes from soybean and cotton by real-time PCR]

[0086] (method)

[0087] Real-time PCR was carried out using the following primer sets and nucleic acid probes using DNA extracted from the same three test samples as the samples used in Example 1 as templates.

[0088] (primer pair)

[0089] For the soybean-derived DREB gene, a primer pair consisting of a nucleic acid having the nucleotide sequences described in SEQ ID NOs: 1 and 2 used in Example 1 was used.

[0090] For the DREB gene derived from cotton, a primer pair composed of nucleic acids having the nucleotide sequences described in SEQ ID NOs: 6 and 7 used in Example 1 was used.

[0091] (nucleic acid probe)

[0092] In the soybean-derived DREB gene, the nucleic acid probe of SEQ ID NO: 5 was designed for the DNA sequence between the primer pair (SEQ ID NO: 1, 2). The 5' end of the nucleic acid probe was labeled with FAM, and the 3' end was modified with TAMRA as a quencher.

...

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Abstract

The genetically-modified crop detection method is for detecting a genetically-modified crop using a polymerase chain reaction, and comprises: a step for amplifying a nucleic acid having a partial sequence of a soybean-derived DREB gene, by using a nucleic acid in a sample to be tested or a nucleic acid extracted from the sample to be tested as a template, and a primer pair which can be used to specifically amplify the partial sequence of the soybean-derived DREB gene; and a step for detecting or quantifying the amplified nucleic acid. The present invention also provides a reagent kit for detecting or quantifying a genetically-modified crop having an introduced soybean-derived DREB gene using a polymerase chain reaction.

Description

technical field [0001] The invention relates to a method for detecting transgenic crops and a kit for detecting transgenic crops. Background technique [0002] Genetically modified crops developed using genetically modified technology have been used as food. Japan has also approved the import and sale of various genetically modified crops such as corn, soybean, rapeseed, cottonseed (cotton), and potato after safety review. In addition, a labeling system is established for genetically modified crops or foods that use them as raw materials, and the designated genetically modified crops and their processed foods are obliged to carry out the labeling (Non-Patent Documents 1 and 2). [0003] On the other hand, outside of Japan, there are cases where GM crops and non-GM crops are grown in close proximity or used in the same processing plant. Therefore, it is possible to detect recombinant DNA derived from GM crops even in processed foods mixed with trace amounts of GM crops and ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12Q1/6823C12N15/09
CPCC12Q1/68C12N15/09
Inventor 野间聪菊池洋介西真名美宫武圣子
Owner NISSHIN SEIFUN GRP INC
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