CAJ | 학술논문

以猪卵巢组织的总 rNA 为模板，采用 rT-PCr 方法扩增猪 Lhx8基因完整的开发阅读框，将该基因克隆到原核表达载体 pET28a （＋）中，构建了原核重组质粒 pET28a （＋）-Lhx8。将重组质粒转化 rosetta（DE3）菌株，经 IPTG 诱导表达及 SDS-PAGE 检测后，发现1条特异的蛋白表达条带，分子量为37 kD 左右。通过对 IPTG浓度和诱导时间的优化，结果显示融合蛋白的最佳诱导浓度为0.4 mmol/L IPTG，最佳诱导时间为37℃诱导12 h。
이저란소조직적총 rNA 위모판，채용 rT-PCr 방법확증저 Lhx8기인완정적개발열독광，장해기인극륭도원핵표체재체 pET28a （＋）중，구건료원핵중조질립 pET28a （＋）-Lhx8。장중조질립전화 rosetta（DE3）균주，경 IPTG 유도표체급 SDS-PAGE 검측후，발현1조특이적단백표체조대，분자량위37 kD 좌우。통과대 IPTG농도화유도시간적우화，결과현시융합단백적최가유도농도위0.4 mmol/L IPTG，최가유도시간위37℃유도12 h。
Using total rNA of pig ovary as PCr template,the full open reading frame (OrF)of pig Lhx8 gene was obtained by rT-PCr and then cloned into pET28a (+)vector.The pET28a (+)-Lhx8 vector was con-structed and confirmed by restriction enzyme digestion and sequence analysis.The recombinant plasmids were transformed into rosetta(DE3)cells and induced by IPTG.The recombinant fusion protein with MW 37 kD was detected by SDS-PAGE.The recombinant fusion protein reached the peak expression after the transformants cul-tured for 12 h at 37 ℃.The optimal concentration of IPTG was 0.4 mmol/L.