鸡肾癌nov基因亚克隆及其表达

SUBCLONING AND EXPRESSION OF THE CHICKEN NEPHROBLASTOMA OVEREXPRESSED GENE

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.