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Polynucleotide, method and kit used for detecting transgenic crops

A polynucleotide and kit technology, which is applied in the fields of methods and kits, polynucleotides, and genetically modified crops detection, can solve the problems of easy missed detection, complicated preparation methods, and large workload.

Pending Publication Date: 2016-08-17
SHANGHAI INST OF MEASUREMENT & TESTING TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, for the detection of multiple components in processed food and feed, such as corn, soybean, potato and other components or the detection of genetically modified products with multiple different strains of a component, the detection work is very complicated
For the detection of such multi-component and multi-strain transgenic products, the use of a single transgenic reference material not only requires a large amount, but also the preparation method is complicated, the workload is heavy, and it is easy to miss the detection

Method used

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  • Polynucleotide, method and kit used for detecting transgenic crops
  • Polynucleotide, method and kit used for detecting transgenic crops
  • Polynucleotide, method and kit used for detecting transgenic crops

Examples

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Effect test

Embodiment 1

[0117] Example 1 Construction of Plasmid Standard Molecules

[0118] According to the construction process of the transgenic crop universal plasmid DNA standard substance of the present invention:

[0119] a. Using databases (such as Genbank, Shanghai GMO safety testing and evaluation sharing service platform) http: / / www.shgmo.org / welcome.htm ) for bioinformatics analysis, and selected the conservative sequence fragments of the insertion element gene covering the existing transgenic element detection methods: cauliflower mosaic virus CaMV35S promoter (269bp), NOS terminator (256bp), neomycin phosphotransferase gene NPTⅡ ( 831bp), Scrophulariaceae mosaic virus FMV 35S promoter (523bp).

[0120] b. Connect the treated sequences in a single copy in a certain order. In this embodiment, the connection sequence is cauliflower mosaic virus CaMV35S promoter (269bp)-NOS terminator (256bp)-neomycin phosphotransferase gene NPTII (831bp )-Scrophulariaceae Mosaic Virus FMV 35S promoter ...

Embodiment 2

[0137] Example 2 Application of Genetically Modified Crops Universal Detection Plasmid Standard Molecules

[0138] The present embodiment provides a method for applying the transgenic crop universal detection plasmid standard molecule, matching primer pair and probe in the transgenic fluorescent PCR detection method, including the following steps:

[0139] The method SNT 1204-2003 "Real-time Fluorescent PCR Qualitative Test Method for Genetically Modified Components in Plants and Its Processed Products" (China Entry-Exit Inspection and Quarantine Industry Standard) was selected for the primer probe and PCR reaction system and reaction time and temperature. As a standard method for transgene detection, the program uses real-time fluorescent quantitative PCR method to simultaneously amplify the exogenous transgenic elements CaMV35S, FMV35S, NOS, NPTII and compare the Ct values ​​obtained by amplifying the standard genes in endogenous plants. When the specific concentration of th...

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Abstract

The invention discloses a polynucleotide, method and kit for detecting transgenic crops. Specifically, the present invention discloses the sequences of the polynucleotide construct pLW10 and its matching primer probes that can be used as standard molecules for real-time fluorescent PCR detection of transgenic crops. The plasmid standard molecule of the present invention solves the problem of lack of standard substances in the real-time fluorescent PCR detection of transgenic crops, ensures the comparability of the detection results of the real-time fluorescent PCR method of the transgenic crops, and provides a reliable quality control method for the real-time fluorescent PCR detection of the transgenic crops.

Description

technical field [0001] The invention relates to a set of plasmid molecules in the technical field of bioengineering, in particular to a set of polynucleotides, methods and kits for detection of transgenic crops. Background technique [0002] Transgenic technology transfers genes between different species, transforms the genetic material of organisms, and transforms them to the goals required by human beings in terms of traits, nutritional quality, and consumption quality, so as to meet various human needs, such as increasing production, expressing special Nutrients even gain resistance to herbicides, viruses or pests. However, with the widespread planting of genetically modified crops around the world, their food safety and environmental safety issues have also attracted the attention of consumers, governments and related agencies. The labeling of genetically modified products has become an important part of the supervision of genetically modified products in various countr...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12N15/63C12Q1/68
CPCC12Q1/6895C12Q2600/166
Inventor 梁文刘刚许丽李妍闻艳丽李兰英任淑贞
Owner SHANGHAI INST OF MEASUREMENT & TESTING TECH
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