分离和鉴定差异表达基因是了解各项生命活动和疾病分子调控机制的重要手段。大多数基因克隆方法不能获得稀少转录的低丰度差异基因 ,且在分离基因的鉴定上易出现假阳性。将抑制消减杂交技术与消减 c DNA文库的高通量筛选结合起来 ,可以...分离和鉴定差异表达基因是了解各项生命活动和疾病分子调控机制的重要手段。大多数基因克隆方法不能获得稀少转录的低丰度差异基因 ,且在分离基因的鉴定上易出现假阳性。将抑制消减杂交技术与消减 c DNA文库的高通量筛选结合起来 ,可以消除各种假阳性和假阴性 。展开更多
Objective: The aim of this study was to use lung cancer targeting binding polypeptide ZS-9 to screen cDNA library of human lung cancer and obtain ZS-9 specific ligand to confirm tumor marker of non small-cell lung can...Objective: The aim of this study was to use lung cancer targeting binding polypeptide ZS-9 to screen cDNA library of human lung cancer and obtain ZS-9 specific ligand to confirm tumor marker of non small-cell lung cancer. Methods: Artificially synthesize biotin labeled peptide ZS-9, anchored ZS-9 in the enzyme label plate coupled by avidin, used ZS-9 as probe to screen cDNA library of human lung cancer, after screening, obtained bacteriophage clone specifically binding with anchored polypeptide ZS-9. Extracted plasmid of bacteriophage and performed sequencing after amplified by PCR. Results: It was demonstrated by bioinformatic analysis on the sequence of ligand binded by lung cancer specific peptide ZS-9 that the ligand was the cytoskeletal protein periplakin on the surface of lung cancer cells, suggesting that periplakin might be a new marker for non-small-cell lung cancer in lung cancer. Conclusion: Use specific lung cancer binding peptide to screen new tumor marker periplakin in lung cancer and further studies on its biologic functions in genesis and development of lung cancer are still needed.展开更多
基金Supported by grants from the Science and Technology Planning Project of Guangdong Province (No. 2010B031600066 No. 2010B031500034+2 种基金 No. 2008B030303008)the Key Scientific Subject Foundation of Ministry of Education of China (No. 208105)the National Science and Technology Major Projects for New Drugs (No. 2011zx09102-001-31)
文摘Objective: The aim of this study was to use lung cancer targeting binding polypeptide ZS-9 to screen cDNA library of human lung cancer and obtain ZS-9 specific ligand to confirm tumor marker of non small-cell lung cancer. Methods: Artificially synthesize biotin labeled peptide ZS-9, anchored ZS-9 in the enzyme label plate coupled by avidin, used ZS-9 as probe to screen cDNA library of human lung cancer, after screening, obtained bacteriophage clone specifically binding with anchored polypeptide ZS-9. Extracted plasmid of bacteriophage and performed sequencing after amplified by PCR. Results: It was demonstrated by bioinformatic analysis on the sequence of ligand binded by lung cancer specific peptide ZS-9 that the ligand was the cytoskeletal protein periplakin on the surface of lung cancer cells, suggesting that periplakin might be a new marker for non-small-cell lung cancer in lung cancer. Conclusion: Use specific lung cancer binding peptide to screen new tumor marker periplakin in lung cancer and further studies on its biologic functions in genesis and development of lung cancer are still needed.
