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Transgenic soybean BPS-CV127-9 transformation event exogenous insertion carrier flanking gene and application thereof

A technology for transgenic soybeans and transformation events, which is applied to the lateral gene of the transgenic soybean BPS-CV127-9 transformation event and its application field. The effect of sensitive method and broad market prospect

Inactive Publication Date: 2012-07-18
INST OF PLANT PROTECTION SHANDONG ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0014] After searching the existing patents and other literatures, we have not found any reports about the foreign insertion of the flanking sequence in transgenic soybean BPS-CV127-9 and the use of this sequence to establish transformant-specific qualitative and quantitative PCR detection

Method used

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  • Transgenic soybean BPS-CV127-9 transformation event exogenous insertion carrier flanking gene and application thereof
  • Transgenic soybean BPS-CV127-9 transformation event exogenous insertion carrier flanking gene and application thereof
  • Transgenic soybean BPS-CV127-9 transformation event exogenous insertion carrier flanking gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0065] Example 1 PCR amplification of transgenic soybean BPS-CV127-9 transformant exogenous insertion sequence

[0066] Extract the genomic DNA of transgenic soybean BPS-CV127-9: use 4 kinds of restriction endonucleases DraI, EcoRV, PvuII, StuI to fully digest, and purify the digested products.

[0067] The synthetic primer sequences are as follows:

[0068] AT1: 5'-CCAACGATGATGTTGTTGATTGGGATG-3';

[0069] AT2: 5'-AACGAATCCCATCACCACAAATCCAAT-3'.

[0070] According to the method provided by the clontech GenomeWalker kit, the adapter primers AP1 and AP2, and the BPS-CV127-9 insertion vector structural sequence were used to design primers for the experiment.

[0071] Using transgenic soybean BPS-CV127-9 genome digestion products as templates, two rounds of PCR amplification were performed. The primer pair AP1 and AT1 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...

Embodiment 2

[0073] Example 2 Application of Transgenic Soybean BPS-CV127-9 Transformation Event of Foreign Source Insertion Vector Side Gene

[0074] 1) Utilize the sequence provided in the present invention to design the transgenic soybean BPS-CV127-9 transformation event transformant-specific qualitative PCR detection method:

[0075] The primers were synthesized by Shanghai Boshang Biological Co., Ltd. The sequences of the synthesized primers are as follows:

[0076] CV127-F: 5'-GCCCTCCTTATTTATCCCCTTAGT-3';

[0077] CV127-R: 5'-AGGGTTTCAGCAGGTTCGTT-3';

[0078] BPS-CV127-9, Conquista, transgenic soybean GTS40-3-2, Mon87701, A5547-127, A5547, transgenic soybean 305423, transgenic corn Mon810, non-transgenic corn Zhengdan 958, transgenic cotton MON531, non-transgenic soybean 1138- The 2 genomes were used as templates, and the CV127-F / CV127-R primer sets were used for PCR amplification. The PCR program was: 95°C 5min pre-denaturation; 94°C 30s, 56°C 30s, 72°C 30s, 35 cycles; 72°C exten...

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Abstract

The invention discloses a transgenic soybean BPS-CV127-9 transformation event exogenous insertion carrier flanking gene, the sequence of which is shown in a sequence table 1. The transgenic soybean BPS-CV127-9 transformation event exogenous insertion carrier flanking gene can be used for specifically and qualitatively detecting whether soybean and borderline products contain a BPS-CV127-9 transformation event, and quantitatively detecting whether the soybean and borderline products contain the BPS-CV127-9 transformation event. According to the invention, the flanking sequence of the transgenic soybean BPS-CV127-9 transformation event exogenous insertion carrier sequence is sequenced and disclosed for the first time; and a specifically qualitative and quantitative PCR (Polymerase Chain Reaction) detection method of the transgenic soybean BPS-CV127-9 transformant is built, and the method is sensitive, accurate, simple and reliable and has broad market prospects.

Description

technical field [0001] The present invention relates to transgenic soybean BPS-CV127-9 transformation event exogenous insertion vector side gene and application thereof. Background technique [0002] With the mass planting and application of genetically modified crops, genetically modified products have begun to enter our lives in large numbers. The potential food safety and ecological safety issues of genetically modified products have attracted people's attention day by day, and countries have formulated corresponding laws and regulations to supervise genetically modified products. Many countries require the labeling of genetically modified products by legislation or otherwise. Except for the United States, Canada, Argentina and Hong Kong Special Administrative Region of China, which implement voluntary labeling systems, most countries in the world, such as EU countries, Japan, South Korea and other countries, have formulated a series of mandatory labeling standards. In ...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12N15/10C12Q1/68
Inventor 路兴波孙红炜李宁李凡杨淑珂
Owner INST OF PLANT PROTECTION SHANDONG ACAD OF AGRI SCI
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