摘要
从东亚三角涡虫cDNA文库中挑选出克隆M455,经BLASTP确定隶属于14-3-3ε基因家族,具有14-3-3蛋白的保守结构域命名为Dj14-3-3ε基因.该基因含有完整的最大开放阅读框(ORF),编码蛋白约10.7 kDa.构建pET28b-Dj14-3-3ε原核表达载体,通过IPTG诱导在大肠杆菌中表达,western blot鉴定,表明原核表达载体构建成功,并在大肠杆菌中正常表达.
M455, which is belonged to 14-3-3 gene family confirmed by BLASTP, was isolated from the eDNA library of Dugesia japonica. We named it Dj14-3-3ε gene as its encoded protein contained a conserved domain of 14-3-3 protein. The largest ORF of its eDNA consisted of 291bp and encoded a protein of 97 amino acids. In this study we construct Prokaryotic expression vector of pET28b-Dj14-3-3,which was induced by TPTG in the E. coli and identified using western blot. The results show that an prokaryotie expression recombinant plasmid pET28b-Dj14-3-3ε has been successfully constructed and expressed in the E. coli.
出处
《山东理工大学学报（自然科学版）》
CAS
2010年第6期104-106,共3页
Journal of Shandong University of Technology:Natural Science Edition
基金
山东省自然科学基金资助项目(ZR2009DM029)
