一个富含脯氨酸肝癌新基因的克隆和抗体制备

The cloning of a new liver cancer related gene and preparation of the antibody

目的 克隆新的肝癌相关基因 ,探索肝癌发生的分子机制。方法 采用DDPCR技术 ,分析原发性肝癌和癌旁肝组织间基因表达差异情况 ,获得差异表达基因片段 ,Northern杂交证实该基因片段能与原标本印迹重现 ;用此基因片段作为探针 ,通过筛选胎盘cDNA文库而获得新基因全长 ;利用GST融合蛋白制备新基因C端多克隆抗体 ,Western杂交验证抗体的特异性和目的基因的表达。结果 成功克隆了一个富含脯氨酸的肝癌新基因cDNA全长 ,并制备出新基因多克隆抗体。结论 我们克隆的这一新的肝癌差异表达基因和制备的该基因C端多克隆抗体为肝癌的基因治疗打下了基础。

Objective To clone a new liver cancer associated gene, and to explore the molecular basis of liver cancer genesis. Methods Using mRNA differential display polymerase chain reaction (DDPCR), we investigated the difference in mRNA expression between primary hepatocellular carcinoma and paracarcinoma liver tissue, and achieved a gene probe. The target gene expression was proved in parent tissue by Northern blot. Screening the placenta cDNA library, we got a full length cDNA of this gene. By GST fusion protein methods, we prepared the special polyantibody of C end of the gene. We identified the polyantibody and the expression of this gene by Western blot method. Results We obtained a full length cDNA of proline riched cancer associated gene, and we have prepared the specific polyantibody of this gene. Conclusion The obtaining of a novel liver cancer associated gene and successful preparation of the polyantibody pave the way for further study on gene therapy for liver cancer.