Flanking sequence of drought-resistant transgenic soybean event FvC5SD-L05 exogenous insertion element and application

A transgenic soybean, fvc5sd-l05 technology, applied in the field of plant biology, can solve the problem that the flanking sequence of the exogenous insert fragment of drought-resistant transgenic soybean has not been found yet

Pending Publication Date: 2021-11-23
JILIN ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] According to the analysis of existing patents and documents, no literature or patent reports related to the flanking sequence of the foreign insert fragment of the drought-resistant transgenic soybean event FvC5SD-L05 have been found.

Method used

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  • Flanking sequence of drought-resistant transgenic soybean event FvC5SD-L05 exogenous insertion element and application
  • Flanking sequence of drought-resistant transgenic soybean event FvC5SD-L05 exogenous insertion element and application
  • Flanking sequence of drought-resistant transgenic soybean event FvC5SD-L05 exogenous insertion element and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1. Analysis of the insertion site of the foreign fragment of the transgenic soybean event FvC5SD-L05

[0042] 1. Extraction of Genomic DNA from Transgenic Soybean FvC5SD-L05

[0043] Weigh 0.1g of soybean sample material, after fully grinding, add 700μl SDS extract, mix well, bathe in 60℃ water for 40min-1h (take out the leaf material after it turns brown), invert and mix once every 10min. Add an equal volume of phenol / chloroform (1:1) and gently invert and mix for 10 minutes, let stand for more than 30 minutes, centrifuge at 10 000 g for 10 minutes, and take the supernatant. Add an equal volume of phenol / chloroform (1:1) and mix well, let stand for 10min. Centrifuge at 10 000 g for 10 min, take the supernatant, add 2 times the volume of absolute ethanol, precipitate, and let stand for more than 10 min. Centrifuge at 10,000g for 1 min, discard the supernatant, and wash the pellet twice with 70% or 75% ethanol. Air dry, add 400μl sterile distilled water to dis...

Embodiment 2

[0047] Example 2. Analysis of the left and right flanking sequences of the foreign insert fragment of the transgenic soybean event FvC5SD-L05

[0048] According to the exogenous insertion sequence of the transgenic soybean event FvC5SD-L05 and the upstream and downstream sequences of the insertion site in the soybean reference genome, PCR detection primers were designed. The primer for amplification of the upstream sequence of the FvC5SD-L05 insertion site is L05-LB-F1

[0049] (5'-TGACCGATAAGCCATCATG-3')

[0050] And L05-LB-R1 (5'-GGTCAACTTCCGTACCGAGC-3'); the downstream sequence amplification primer of FvC5SD-L05 insertion site is L05-RB-F1 (5'-ACGGCTACACTTACCTTGTCCTGA-3')

[0051] and L05-RB-R1 (5'-TTGTCTGGTCTGAAGACCTTTCTA-3').

[0052]Using the FvC5SD-L05 genomic DNA as a template, the above primers were used for PCR amplification. The PCR reaction system (50uL) is: 20×PCR buffer 5uL, 10mmol / L dNTPs 1uL, 5U / uL Taq enzyme 1uL, DNA sample 2.0uL, 10umol / L forward primer 1u...

Embodiment 3

[0054] Example 3. Transgenic soybean event FvC5SD-L05 specific PCR detection

[0055] According to the left border flanking sequence (as shown in SEQ-2) and the right border flanking sequence (as shown in SEQ-3) of the exogenous insert fragment of the transgenic soybean event FvC5SD-L05, specific detection primers were designed respectively. In the left border flanking sequence-specific detection primer combination, one of the primers is a forward primer designed according to the sequence of the 1-197th position of SEQ-2, as shown in SEQ-4; the other primer is based on the 198th- The reverse primer designed for the 1749 site sequence is shown in SEQ-5. In the right border flanking sequence-specific detection primer combination, one of the primers is a forward primer designed based on the sequence of SEQ-3 1-1187, as shown in SEQ-6; the other primer is based on SEQ-3 1188- The reverse primer designed for the 2174 site sequence is shown in SEQ-7.

[0056] DNA samples were extr...

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Abstract

The invention relates to a flanking sequence of a drought-resistant transgenic soybean event FvC5SD-L05 exogenous insertion element and application, which is characterized by comprising a left boundary flanking sequence and a right boundary flanking sequence of the transgenic soybean event FvC5SD-L05 exogenous insertion element; the sequence can be used as a target DNA sequence to establish a specific detection method of the transgenic event. The flanking sequence of the exogenous insertion element and the detection method are suitable for specific detection of transgenic soybean events including parent and derivative strains or varieties and products thereof including plants, tissues, seeds and products.

Description

technical field [0001] The invention relates to a flanking sequence of a drought-resistant transgenic soybean event FvC5SD-L05 exogenous insertion fragment and its application, belonging to the field of plant biotechnology. Background technique [0002] Soybean is native to China and has a long history of cultivation. It is one of the main sources of protein and oil. Abiotic stresses such as drought and salt damage have a greater impact on soybean production. Therefore, carrying out functional research on genes related to soybean drought resistance has great theoretical and practical value for revealing the mechanism of soybean drought resistance and cultivating soybean drought-resistant varieties. The main soybean producing areas in my country are mainly distributed in arid, saline-alkali, and alpine regions. Severe growing environment, especially drought and soil salinization, pose a serious threat to my country's soybean production. According to incomplete statistics, t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/13
Inventor 张玲董英山邱红梅易立
Owner JILIN ACAD OF AGRI SCI
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