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
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
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...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More 


