摘要
采用PCR定点突变技术对睫状神经营养因子(CNTF)基因进行改造,获得了CNTF衍生物(CNTF-D)基因序列分析证明其核苷酸序列符合设计要求,并在大肠杆菌中获得高效表达,CNTF-D表达量超过40%,复性后,经Q-Sepharose HP纯化,其纯度超过95%,体内法测定发现能明显降低实验小鼠的体重。表明CNTF-D在体内具有良好的生物学活性,为下游开发奠定了基础。
Based on the natural human CNTF gene, the mutated hCNTF gene was generated by site-specific mutagenesis. The gene was inserted into E. coli and confirmed by DNA sequencing. The high expression of CNTF-D has been achieved. The expression level of recombinant protein reached to more than 40%. The purity of CNTF-D was about 95% after denatu- ring and purifying by Q-Sepharese HP chromatography. The biological activity of CNTF-D was measured by mouse in vivo test. The results showed that this mutant protein could obviously reduce body weight of tested mouse. It laid a foundation in development of a new type of high efficient body weight reducing drug .
出处
《微生物学免疫学进展》
2006年第4期10-13,共4页
Progress In Microbiology and Immunology
关键词
睫状神经营养因子
点突变
基因表达
Cillary neurotrophic factor （CNTF）
Site-specific mutagenesis
Gene expression
