Utilization of whole genome PCR to identify genes possibly regulated by MTF-1

• Main Authors: Ya-Ting Zhuang, 莊雅婷
• Other Authors: Lih-Yuan Lin
• Format: Others
• Language: zh-TW
• Published: 2006
• Online Access: http://ndltd.ncl.edu.tw/handle/59000550330549335371

碩士 === 國立清華大學 === 分子與細胞生物研究所 === 94 === The transcriptional activator metal-responsive transcription factor (MTF-1) exerts its function by activating down-stream genes to maintain heavy metal homeostasis and to respond to various oxidative stresses. A limited genes have been found recently that are regulated by MTF-1. In this study, we tried to employ whole genome PCR to search unidentifiied target genes of MTF-1. DNA fragment recovered after screening have been cloned, and sequenced. Several DNA fragments that contain MREs (metal response element) were obtained. Using EMSA (Electrophoretic Mobility Shift Assay) and reporter gene assay, the MTF-1 binding and transactivation activities of DNA fragment were examined. The DNA sequences were also searched from blast/NCBI human genome database to reveal the identity of the DNA. The sequences locate in the promoter region of several genes, such as zinc finger protein 268, brain-specific angiogenesis inhibitor 1. Since the sequences present far away from the identified genes, we speculate that the DNA sequence may serve as enhancer of the genes regulated by MTF-1.