摘要
为构建pET-32a-2SS28-SS14和pET-32a-3SS28原核表达质粒并检测其在大肠杆菌中的表达,将两种形式的生长抑制素SS14和SS28基因克隆到pET-32a载体中;酶切及测序鉴定后,采用不同终浓度的IPTG在37℃条件下进行诱导表达。Western blot结果显示:两组分别诱导出分子量大小为28.4 kD和29.8 kD的目的蛋白,不同终浓度的IPTG诱导对目的蛋白的表达量没有明显影响。目的蛋白主要为可溶性蛋白,少数以包涵体形式存在。本研究为生长抑制素基因工程疫苗生产应用奠定了基础。
In order to construct pET-32a-2SS28-SS14 and pET-32a-3SS28 prokaryotic expression plasmids and detect their expression in E.coil,both SS14 and 5S28 somatostatin genes were cloned into pET-32 a vector;and after enzyme digestion and sequencing,the inducible expression of target proteins was conducted at 37 ℃ by using different final concentrations of IPTG.The Western blot showed that the induction at different final concentrations of IPTG had an insignificant effect on the expression of target proteins,and the induced expression molecular weights of both somatostatin genes were 28.4 kD and 29.8 kD respectively.The products were mainly a soluble protein and less inclusion body.The research laid the basis for production and application of somatostatin engineering vaccine.
出处
《上海农业学报》
CSCD
2016年第1期15-18,共4页
Acta Agriculturae Shanghai
