鸡肾癌nov基因亚克隆及其表达

鸡肾癌nov基因亚克隆及其表达 SUBCLONING AND EXPRESSION OF THE CHICKEN NEPHROBLASTOMA OVEREXPRESSED GENE 蒋达和 Jiang Dahe 1 2 first-author 李文鑫 Li Wenxin 1 2 武汉大学生命科学学院 武汉大学生命科学学院 School of Life Science,Wuhan University School of Life Science,Wuhan University 武汉大学生命科学学院,武汉,430072 武汉大学生命科学学院,武汉,430072 采用DNA重组技术将鸡肾癌nov基因的外显子4和部分外显子3、5的cDNA亚克隆于表达载体pET-3d质粒中,并在大肠杆菌BL21中诱导表达。免疫沉淀法和SDS-聚丙烯酰胺凝胶电泳分析表明,nov基因表达的NOV融合蛋白分子量约2.5×10 4 ,与序列推测的分子量基本相符。 An amino-terminal-truncated chicken nov （nephroblastoma overexpressed gene ） cDNA sequence,which contains a complete exon 4 and partial exons 3 and 5,was sub-cloned in the expression vector pET-3d to construct the recombinated plasmid pET-3d Sau 11 clone. The recombinant was induced by IPTG to express an amino-terminal-truncated nov gene product （a fused protein） in E. coli BL 21. Molecular weight of the fused protein was about 2. 5×104 which was determined by SDS-polyacrylamide gel electrophoresis and identi-fied by immunoprecipition test. It was consistent with the molecular weight of the putative polypeptide according to the nov cDNA sequence. nov基因 基因表达 融合蛋白 免疫沉淀 nephroblastoma overexpressed gene gene expression fusion protein im-munoprecipition R737.11 国家自然科学基金资助的课题 1994-06-01 2021-04-01 6