CAJ | 학술논문

通过双酶切、连接转化等方法将人乙醛脱氢酶2（ALDH2）基因克隆至载体 pGEX-4T-1上，构建原核表达重组质粒 pGEX-4T-1-ALDH2。重组质粒转化至 E．coli BL21（DE3）后经 IPTG 诱导，SDS-PAGE 分析表明目的蛋白得到大量表达。由于此载体本身带有融合蛋白标签 GST（谷胱甘肽转移酶），所以得到了部分可溶性的目的蛋白。通过对表达条件进行探讨，发现在28℃下，以终浓度0.1 mmol·L-1的 IPTG 诱导表达10 h，可溶性的目的蛋白约占总蛋白的25％。表明，融合蛋白标签 GST 及较低的诱导温度有利于提高人乙醛脱氢酶2基因在大肠杆菌内的可溶性表达。
통과쌍매절、련접전화등방법장인을철탈경매2（ALDH2）기인극륭지재체 pGEX-4T-1상，구건원핵표체중조질립 pGEX-4T-1-ALDH2。중조질립전화지 E．coli BL21（DE3）후경 IPTG 유도，SDS-PAGE 분석표명목적단백득도대량표체。유우차재체본신대유융합단백표첨 GST（곡광감태전이매），소이득도료부분가용성적목적단백。통과대표체조건진행탐토，발현재28℃하，이종농도0.1 mmol·L-1적 IPTG 유도표체10 h，가용성적목적단백약점총단백적25％。표명，융합단백표첨 GST 급교저적유도온도유리우제고인을철탈경매2기인재대장간균내적가용성표체。
The human acetaldehyde dehydrogenase 2(ALDH2)gene was cloned into prokaryotic expression vector pGEX-4T-1 with fusion protein tag GST to construct prokaryotic expression recombinant plasmid pGEX-4T-1-ALDH2.The recombinant plasmid was transformed into E.coli BL21 (DE3 ).After being induced by IPTG,expression of the purpose protein was verified by SDS-PAGE analysis,and through optimizing the ex-pression conditions(0.1 mmol· L-1 IPTG,28 ℃,10 h),the purpose protein present in supernatant achieved 25% of that in pellet of the bacterial lysate because of the fusion protein tag GST.It showed that the fusion pro-tein tag GST and low temperature could improve soluble expression of ALDH2 in Escherichia coli .