Abstract
The full-length cDNA of the soybean seeds was obtained by RT-PCR, and amplified by self-designed primers to obtain the target gene αc, The target gene was linked to the vector to construct a recombinant cloning vector pGEM-αc, which was identified by double digestion with XhoⅠ/EcoRⅠ and ligated with vector pET-28a to construct a recombinant plasmid pET-28a-αc. After correctly transferred into competent cells E.coli BL21 (DE3), the recombinant protein of α-subunit core region was expressed by inducing with the OD600 nm value 0.8, isopropylthiogalactoside (IPTG) 0.2 mmol/L at 37 ℃ for 9 h, and the molecular weight of the protein was about 50 kV. The cell of engineering bacteria pET-28a-αc-BL21 was ultrasonically broken and centrifuged at low temperature. Target protein in the supernatant was purified with AKTA protein purification system using nickel ion affinity chromatography column.
Publication Date
5-28-2020
First Page
15
Last Page
19,38
DOI
10.13652/j.issn.1003-5788.2020.05.003
Recommended Citation
Yao-xi, LI; Yan-qiu, YUAN; Qian, JU; Meng-nan, GAO; Shu-yi, ZHOU; Ya-yun, HU; and Guang-zhong, LUAN
(2020)
"Cloning,expression and purification of β-conglycin inα-subunit core region,"
Food and Machinery: Vol. 36:
Iss.
5, Article 3.
DOI: 10.13652/j.issn.1003-5788.2020.05.003
Available at:
https://www.ifoodmm.cn/journal/vol36/iss5/3
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