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目的 :分离在肝硬化组织上调表达的新基因 ,以阐述肝硬化发生的分子机制。方法 :利用优化的代表性差异分析方法分析正常肝脏及肝硬化组织表达基因的差异。两轮杂交后的产物克隆到T载体中 ,斑点杂交筛选阳性克隆以获得差异表达基因片段 ,并进一步以半定量RT PCR鉴定。结果 :上调表达的克隆包括与一些序列和功能完全未知的基因同源的cDNA以及未曾报道过的在肝脏病变中发生表达变化的已知基因———钙结合酪氨酸磷酸化调节蛋白、雌激素应答B盒蛋白等。结论 :利用优化的代表性差异分析方法成功分离获得在肝硬化组织中上调表达的基因片段 ,其中一些新基因的克隆及其在肝硬化发生中的作用有待进一步研究。
OBJECTIVE: To isolate new genes that are upregulated in cirrhotic tissues to elucidate the molecular mechanism of cirrhosis. Methods: The difference of expression genes in normal liver and cirrhosis tissues was analyzed by optimized representative difference analysis. After two rounds of hybridization, the cloned products were cloned into T vector, positive clones were screened by dot blot hybridization to obtain differentially expressed genes, and further identified by semi-quantitative RT PCR. Results: The up-regulated clones included cDNAs homologous to some unknown genes and unknown genes that have not been reported in the liver lesions. Calcium-associated tyrosine phosphorylation regulator, Hormone response B box protein and so on. Conclusion: The gene fragments up-regulated in cirrhotic tissues were successfully isolated by the optimized representative differential analysis. Cloning of some new genes and their roles in the development of cirrhosis need further study.