猪繁殖候选基因Lhx8的原核表达及条件优化  

Prokaryotic expressing and optimization of reproduction candidate gene Lhx8 in pigs

作　　者：王晶晶[1] 毛慧[1] 陈琳[1] 何闪[1] 潘雪男[1] 

机构地区：[1]上海市农业科学院畜牧兽医研究所,上海201106

出　　处：《上海农业学报》2015年第4期54-57,共4页Acta Agriculturae Shanghai

摘　　要：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12h。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- strutted and confirmed by restriction enzyme digestion and sequence analysis. The recombinant plasmids were transformed into Rosetta（ DE3 ） ceils 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.

关 键 词：猪 Lhx8基因 原核表达 条件优化 

分 类 号：Q78[生物学—分子生物学]