【摘要】：為獲得高活性重組人乙醛脫氫酶2(ALDH2)重組蛋白,研究通過PCR獲得ALDH2基因,先后構(gòu)建克隆和表達(dá)載體p GEM-ALDH2和p TYB11-ALDH2。轉(zhuǎn)化的陽性重組菌經(jīng)IPTG誘導(dǎo),SDS-PAGE顯示融合蛋白獲得大量表達(dá)。通過正交試驗(yàn)對表達(dá)條件優(yōu)化設(shè)計(jì),結(jié)果表明,誘導(dǎo)OD值0.3,30℃,IPTG濃度1.0 mmol·L-1,誘導(dǎo)表達(dá)4 h為最優(yōu)表達(dá)條件。由于融合蛋白以包涵體形式存在,經(jīng)變性、復(fù)性,使用IMPACTTM-CN蛋白純化系統(tǒng),經(jīng)內(nèi)含肽標(biāo)簽蛋白自裂解洗脫、純化,獲得僅含有幾個載體氨基酸的ALDH2蛋白,蛋白濃度為0.045 mg·m L-1。純化的ALDH2蛋白比活力為462 U·mg-1。
[Abstract]:In order to obtain recombinant human aldehyde dehydrogenase 2 (ALDH2) recombinant protein with high activity, the ALDH2 gene was obtained by PCR. The cloning and expression vectors of p GEM-ALDH2 and p TYB11-ALDH2. were constructed successively. The positive recombinant strain was induced by IPTG, and SDS-PAGE showed that the fusion protein was highly expressed. The expression conditions were optimized by orthogonal experiment. The results showed that the optimal expression conditions were induced OD value of 0.33 鈩，

本文編號：2420687