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Para-gene for exogenous insertion vector of transgenic maize transformation event MON88017 and application thereof

A technology of MON88017 and transgenic corn, which is applied in the direction of DNA preparation, recombinant DNA technology, DNA/RNA fragments, etc., can solve the problems of specific qualitative and quantitative PCR detection of transformants that have not been found yet

Inactive Publication Date: 2011-10-05
INST OF PLANT PROTECTION SHANDONG ACAD OF AGRI SCI
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  • Abstract
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Problems solved by technology

[0013] After searching the existing patents and other literatures, no report has been found on the transgenic maize Mon88017 exogenously inserted flanking sequence and using this sequence to establish transformant-specific qualitative and quantitative PCR detection

Method used

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  • Para-gene for exogenous insertion vector of transgenic maize transformation event MON88017 and application thereof
  • Para-gene for exogenous insertion vector of transgenic maize transformation event MON88017 and application thereof
  • Para-gene for exogenous insertion vector of transgenic maize transformation event MON88017 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0072] Example 1 PCR amplification of the side sequence of the transgenic maize Mon88017 transformant inserted into the vector

[0073] The genomic DNA of transgenic maize Mon88017 was extracted, and the genomic DNA of Mon88017 was digested according to the method provided by the kit using the BD GenomeWalker kit from Clontech, USA.

[0074] The synthetic primer sequences are as follows:

[0075] P-ractgsp1: 5'-CTTTAGGACTTTAGGGGTTGTT-3';

[0076] P-ractgsp2: 5'-GGACTATCCCGACTCTCTTC-3'.

[0077] The transgenic maize MON88017 was used as a template for two rounds of PCR amplification, the primer pair AP1 and P-ractgsp1 were used for the first round of PCR reaction, and the amplification system was 20 μL. The PCR program was 94°C for 25s, 72°C for 3min, 6 cycles; 94°C for 25s, 64°C for 30s, 36 cycles; 64°C for 7min, 1 cycle. The primer pair AP2 and P-ractgsp2 were used for the second round of PCR, in which 1 μL of the 50-fold dilution of the first round of amplification produc...

Embodiment 2

[0079] Example 2 Application of side gene of transgenic maize Mon88017 transformation event of exogenous insertion vector

[0080] 1) Using the sequence provided in the present invention to design a transformant-specific qualitative PCR detection method for the transformation event of transgenic maize Mon88017:

[0081] The primers were synthesized by Shanghai Boya Biological Company, and the primers were diluted to 10 μmol / L for use. The synthetic primer sequences are as follows:

[0082] Mon88017-F: 5'-TCCTGAACCCCTAAAATCCC-3';

[0083] Mon88017-R: 5'-TTTCTCATCTAAGCCCCCAT-3'.

[0084] Nine kinds of transgenic corn MON88017, MON810, MON863, NK603, T25, TC1507, Bt11, Bt176, GA21 were extracted, non-transgenic corn Zhengdan 958, transgenic cotton MON531, non-transgenic cotton Zhong49, transgenic soybean GTS40-3-2, Non-transgenic soybean 1138-2 gene DNA was used as a template, and PCR amplification was carried out by using the primer combination of Mon88017-F and Mon88017-R re...

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Abstract

The present invention discloses a para-gene for an exogenous insertion vector of transgenic maize transformation event MON88017 and application thereof, the sequence of which is shown in table 1. The para-gene for the exogenous insertion vector of transgenic maize transformation event MON88017 provided by the invention can be used for specifically detecting whether the transgenic maize and related products contain transformation event MON88017 and the content of the transformation event; and the detection method is sensitive, accurate, simple and reliable, and has broad market prospects. The para-gene analyzes and confirms the sources of different bases in the para-sequence of the exogenous insertion vector of transgenic maize transformation event MON88017 for the first time, and determines the binding sites of the exogenous insertion vector sequence and the maize genome sequence, thus contributing to scientific research.

Description

technical field [0001] The invention relates to a side gene of a transgenic maize MON88017 transformation event exogenously inserted into a vector and application thereof. Background technique [0002] In recent years, genetically modified crops such as corn, soybean, cotton, rapeseed and tomato have been approved for planting and production in many countries, and some have been processed into food, feed or used as food additives. Since the ecological safety and food safety of genetically modified products have been controversial, more than 40 countries and regions have successively implemented genetically modified product labeling systems. [0003] The establishment of genetically modified labeling systems in various countries has put forward strict requirements on the sensitivity and accuracy of genetically modified detection technology, and various genetically modified detection technologies have also become research hotspots. Commonly used transgenic detection methods i...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12N15/10C12Q1/68
Inventor 路兴波袁磊孙红炜武海斌李凡韩伟
Owner INST OF PLANT PROTECTION SHANDONG ACAD OF AGRI SCI
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