期刊论文

中文

中国生物工程杂志

1671-8135

2008

28

3

53-58

10.3969/j.issn.1671-8135.2008.03.009

动物营养与饲料科学湖北省重点实验室开放基金（2006A008）； 湖北省自然科学基金（2005ABA094）资助项目；

本校为第一机构

根据已报道的单链Monellin甜蛋白的氨基酸序列,按大肠杆菌基因偏爱密码子,设计和人工合成了单链monellin基因。将单链monellin基因克隆到大肠杆菌表达载体pET-28a中,构建了重组表达载体pET28a-mon,转化大肠杆菌BL21（DE3）,得到表达Monellin的大肠杆菌工程菌株。借助SDS-PAGE分析方法,研究了乳糖代替IPTG诱导大肠杆菌表达甜蛋白Monellin。通过对乳糖作为诱导剂表达条件进行优化,Monellin的表达量可占细胞总蛋白的33.09%,与IPTG诱导表达量接近。为乳糖作为诱导剂应用于重组大肠杆菌生产甜蛋白Monellin提供了参考依据。

According to the amino acid sequence of Monellin and the bias in coden choice in Escheria coli, a single chain monellin gene was synthesized and subcloned into a Escheria coli expression vector pET-28a. The recombined plasmid pET28a-mon was then transformed into Escheria coli BI21 （ DE3 ）. Lactose was used as an inducer instead of IPTG. Through proper optimization of induction conditions, an expression level 33.09% of total cellular protein was achieved. The expression level of recombinant protein induced by lactose was basically the same as ...