Application of soy protein
A technology of soybean protein and protein, applied in the field of bioengineering, can solve problems such as the pollution risk of protein biological products, and achieve good results
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
preparation example Construction
[0020] Further, the preparation method of the soybean protein comprises the following steps:
[0021] S01: amplifying the coding sequence of the soybean protein;
[0022] S02: constructing a recombinant plasmid comprising the coding sequence;
[0023] S03: transforming the recombinant plasmid into competent cells, and inducing expression;
[0024] S04: Separating and purifying the bacterial liquid after the induced expression to obtain the soybean protein.
[0025] Further, in the above step S01, the primer pair for amplifying the coding sequence of the soybean protein is as shown in SEQ ID NO:2 and SEQ ID NO:3:
[0026] SEQ ID NO:2: 5'-CGCCCATATGGCCAGCGTTGAGGTTGC(Nde I)-3',
[0027] SEQ ID NO: 3: 5'-ACCGCTCGAGCTAAGCTTCAGTCTTCTCAAC(Xho I)-3'.
[0028] According to the sequence characteristics of the PMD19-T carrier and the target gene, the above-mentioned primer pair introduces the homologous sequence of the corresponding enzyme cutting site, and introduces the homologous ...
Embodiment 1
[0035] The cloning of embodiment 1 gene
[0036] According to the characteristics of the PMD19-T carrier and gene sequence, the homologous sequence of the corresponding enzyme cutting site was introduced, and two endonucleases (Nde I) and (Xho I) were respectively introduced into the 5' ends of the upstream and downstream primers of the amplified soybean protein gene. end homologous sequences.
[0037] Upstream primer F: 5'-CGCCCATATGGCCAGCGTTGAGGTTGC-3',
[0038] Downstream primer R: 5'-ACCGCTCGAGCTAAGCTTCAGTCTTCTCAAC-3'.
[0039] The upstream primer F and the downstream primer R were used as a primer pair, and the soybean R0 radicle cDNA was used as a template for PCR amplification. The reaction system was 4 μL of 5×Phusion HF Buffer, 1.6 μL of 2.5 mM dNTP, 10 mM of upstream and downstream primers, 1 μL of cDNA template, 0.6 μL of DMSO, and made up to 20 μL with water. Pre-react at 98°C for 30s and react at 98°C for 5s on a PCR instrument. The annealing temperature depend...
Embodiment 2
[0041] Preparation and transformation of embodiment 2 supercompetent
[0042] Prepare Escherichia coli TOP10 and BL21Star competent according to the method provided by the super-competent kit provided by Shanghai Sangong, pick the streak-cultured Escherichia coli monoclonal strain in 5 mL of LB liquid medium in the ultra-clean work bench, ℃, 180rmp constant temperature shaker for 12h, inoculated into 50mL freshly prepared LB medium according to the ratio of 1:50, to OD 600 The value is about 0.6. Collect the bacteria in a 50mL centrifuge tube and centrifuge at 8,000g for 3min. After discarding the supernatant, place the bacteria on ice for 10 minutes, centrifuge at 5,000 rpm at 4°C for 5 minutes, resuspend the bacteria with Buffer A provided in the kit, place on ice for 30 minutes, centrifuge at 5,000 rpm at 4°C for 5 minutes, discard the supernatant , resuspend the bacteria with Buffer B provided in the kit (pre-cooled at -20°C before use), and dispense into sterilized 1.5m...
PUM
| Property | Measurement | Unit |
|---|---|---|
| Column length | aaaaa | aaaaa |
| Diameter | aaaaa | aaaaa |
Abstract
Description
Claims
Application Information
Login to View More