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笔者曾通过分子杂交、Run-off转录活性的检测及原位杂交技术证明,肝癌细胞中N-ras癌基因的表达明显高于正常肝细胞,其分子机理可能是由于转录水平的异常.进而采用Southwestern Blot技术对反式作用因子(trans-acting factor)进行检测,结果发现在肝癌细胞中缺失了一组(分子量45 000、40 000和35 000)DNA结合蛋白.本实验对该组DNA结合蛋白进行分离、鉴定并利用体外转录系统证明其具有抑制转录的作用,提示存在N-ras癌基因的负调控因子.
The authors have demonstrated through molecular hybridization, Run-off transcriptional activity detection and in situ hybridization that the expression of N-ras oncogene in hepatocellular carcinoma cells is significantly higher than that in normal hepatocytes, and the molecular mechanism may be due to the abnormal transcriptional level. Southwestern Blot technology to trans-acting factor (trans-acting factor) was detected in the liver cancer cells found in a group (molecular weight 45 000,40 000 and 35 000) DNA binding protein.In this experiment, the group of DNA binding protein Isolation, identification and use of in vitro transcription system to prove that it has the role of inhibition of transcription, suggesting the presence of N-ras oncogene negative regulatory factor.