羊生长激素在大肠杆菌中的表达及分泌初探

EXPRESSION AND SECRETION OF OVINE GROWTH HORMONE IN E.COLI

利用PCR方法,将羊生长激素(oGH)成熟蛋白编码序列扩增,再将扩增产物接入分泌型表达载体pET12a(SalI位点)中,利用宿主菌自身的OmpT信号肽引导羊生长激素在E.coliBL21(DE3)中分泌,获得正向插入重组质粒pET12-oGH8,对重组子及对照菌进行诱导表达,薄层色谱扫描(SDS-PAGE)检测结果发现,oGH在OmpT信号肽引导下未见明显分泌,推测oGH自身的氨基酸序列可能是影响其在E.

With PCR method, ovine Growth Hormone (oGH) mature protein coding sequence was amplified and fused at the Sal Ⅰ site of secretion vector pET12a, which makes oGH gene under the control of OmpT signal peptide sequence of E.coli BL21(DE3). A desired recombinant pET12 oGH8 was obtained. After induced by IPTG, oGH was expressed in E.coli , but did not detect the secretion. It is assumed that the amino acid sequence of oGH is possibly the factor that influence the secretion in E.coli .