计算机辅助设计改善重组人神经珠蛋白(hNGB)在E.coli中的表达

人神经珠蛋白(hNGB)是新发现的人类脑特异的携氧蛋白.为对其结构和功能进行深入的研究,首先,利用分子生物学实验辅助设计软件BioSun1.0分析了hNGB基因与pBV221载体连接处的二级结构和hNGB基因的局部密码子偏爱性,并依据原核系统外源基因高效表达数学模型对hNGB基因进行了高效表达设计;其次,构建了hNGB基因编码区原核表达载体pBV221-hNGB;最后通过转化大肠杆菌JM109,热激诱导,获得了hNGB基因的高效表达.表达水平约为12%,从而为进一步研究hNGB基因的结构和功能奠定了重要基础.

Improving of Expression of Human Neuroglobin In The E.coli By the Assistant Design of BioSun 1.0

Human neuroglobin (hNGB) is a recently discovered specific oxygen-binding respiratory protein in human brain. In order to further research its structure and function, hNGB gene was subcloned into express vector pBV221,which analyzed by the BioSun 1.0, then the pBV221-hNGB was transformed into E.coli JM109.It was induced at 42 ℃,then subsequent SDS-PAGE electrophoresis shows that pBV221-hNGB led to overexpression of hNGB. We have established an important foundation for further investigation of relationship between structure and function of hNGB.