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The right flank sequence of exogenous insertion vector of a transgenic soybean wh8013 transformation event

A technology of WH8013, transgenic soybeans, applied in biochemical equipment and methods, measurement/inspection of microorganisms, DNA/RNA fragments, etc., can solve problems such as obtaining HAL1 transgenic soybeans that have not yet been researched, lack of detection methods for HAL1 transgenic soybeans, etc., and achieve Good agronomic traits, good specificity, and the effect of improving identification efficiency

Inactive Publication Date: 2020-04-21
JILIN ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are already some HAL1 transgenic plants, but HAL1 transgenic soybeans have not been researched, and there is still a lack of methods that can effectively detect HAL1 transgenic soybeans

Method used

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  • The right flank sequence of exogenous insertion vector of a transgenic soybean wh8013 transformation event
  • The right flank sequence of exogenous insertion vector of a transgenic soybean wh8013 transformation event
  • The right flank sequence of exogenous insertion vector of a transgenic soybean wh8013 transformation event

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1 Obtaining the Right Wing Sequence of Transgenic Soybean WH8013 Insertion Vector

[0046] 1. Experimental materials

[0047] Transgenic soybean WH8013 (provided by the Soybean Research Group of the Institute of Biology, Jilin Academy of Agricultural Sciences): soybean Williams 82 was used as the recipient material, and the plasmid pCHAL1 was used as the transformation vector. WH8013 transformation event) was obtained, and the map of the plasmid pCHAL1 is shown in the attached drawing of the description figure 1 The transgenic soybean WH8013 exhibits salt-alkali tolerance and good agronomic traits, and the insertion site of its T-DNA is finally determined as nucleotide 21177109 on chromosome 8 of soybean through Example 1.

[0048] 2. Acquisition of the right flank sequence

[0049] 1. Extraction of DNA

[0050] (1) Sample collection: The leaves of the transgenic soybean WH8013 planted in the field were collected in mid-June and stored at -80°C.

[0051] (2)...

Embodiment 2

[0063] The specific PCR detection of embodiment 2 transgenic soybean WH8013

[0064] 1. Design of PCR primers

[0065] According to the right border flanking sequence and T-DNA sequence of the transgenic soybean WH8013 obtained in Example 1, a primer pair capable of specifically detecting T-DNA insertion events was designed.

[0066] A primer of the primer pair is on the T-DNA, and the other primer is on the soybean genome: as the 3'F upstream primer is on the T-DNA, the 3'R downstream primer is on the right border genome ( Figure 5 ). The primer pair 3'F (SEQ ID NO: 6) and 3R' (SEQ ID NO: 7) sequences are shown in Table 2.

[0067] Table 2 Primer pairs for specific PCR detection of transgenic soybean WH8013

[0068]

[0069] 2. Specific PCR detection of transgenic soybean WH8013

[0070] Using the genomic DNA of transgenic soybean WH8013 as a template, PCR detection was performed using the above-mentioned PCR primer pair. Wherein, the specific reaction conditions are...

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Abstract

The invention relates to a right flank sequence of an exogenous insertion vector of a transgenic soybean WH8013 transformation event, application of the right flank sequence, a primer pair, a PCR kit and a method for using the primer pair and the PCR kit. Wherein, the nucleotide sequence of the right flank sequence of the present invention is shown in SEQ ID NO: 4, which can specifically indicate the transgene inserted into the T-DNA fragment of pCHAL1 at the 21177109th nucleotide of chromosome 8 soybeans. For the first time, the present invention has published the right flank sequence of the exogenous insertion vector of the transformation event of transgenic soybean WH8013, and established a specific qualitative PCR detection method for transgenic soybean WH8013. The method is sensitive, accurate, simple, reliable and has broad market prospects.

Description

technical field [0001] The invention relates to the field of transgenic soybeans, in particular to a right flank sequence of an exogenous insertion vector of a transgenic soybean WH8013 transformation event. Background technique [0002] Soybean is the main oil crop in the world. In 2015, the global planting area of ​​genetically modified crops reached 179.7 million hectares, of which soybean planting area was 91.6 million hectares, accounting for 51% of the total area [1] . China is not only a big agricultural producer, but also a big agricultural consumer. Since 1996, China has become a net importer of soybeans. With the continuous increase of domestic demand, the import of genetically modified soybeans is also increasing year by year. In 2015 alone, China imported 81.69 million tons of genetically modified soybeans, a year-on-year increase of 14.4%. [2] . [0003] Scientific research and evaluation on the biosafety of genetically modified soybeans is an important te...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/11C12Q1/6895
CPCC12Q1/6895C12Q2600/13
Inventor 于志晶尚丽霞蔡勤安马瑞庄丹任鹏飞王丹
Owner JILIN ACAD OF AGRI SCI