Cloning,expression and purification of β-conglycin inα-subunit core region
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    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.

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李垚熹,袁艳秋,巨倩,等.β-伴大豆球蛋白α-亚基核心区的基因克隆、表达及纯化[J].食品与机械英文版,2020,(5):15-19,38.

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  • Online: February 15,2023
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