The promoter elements of the mouse myelin basic protein gene function efficiently in NG108-15 neuronal/glial cells.
We measured transiently-expressed beta-galactosidase activity by introducing the mouse myelin basic protein (MBP)-lacZ chimeric gene (MBP-lacZ) into the NG108-15 neuronal/glial hybrid cell line. Deletion studies of the promoter region of the MBP gene showed that the promoter region between -1318 bp and -254 bp might contain sequences that repress MBP promoter activity. Fine deletion analysis using BAL 31 exonuclease revealed sequences between bp -208 and -140, -139 and -118, and -89 and -75 which were critical for promoter activity in NG 108-15 cells. DNaseI footprinting analysis revealed a cellular factor(s) that bind to the promoter region between bp -127 and -106 with NG108-15 whole cell extracts. The SV40 promoter was activated by insertion of the sequences around the region protected in footprinting experiments, in a manner independent of its orientation in NG108-15 cells. This protected region is thought to be one of the critical cis-acting DNA elements for efficient transcription.
Miura
M
M
Department of Biological Regulation, National Institute for Basic Biology, Okazaki, Japan.
Tamura
T
T
Aoyama
A
A
Mikoshiba
K
K
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D002678:Chimera; D002872:Chromosome Deletion; D003850:Deoxyribonuclease I / Q000378:metabolism; D006822:Hybrid Cells; D051379:Mice; D008969:Molecular Sequence Data; D004676:Myelin Basic Protein / Q000235:genetics*; D011401:Promoter Regions, Genetic; D013539:Simian virus 40 / Q000235:genetics; D014158:Transcription, Genetic; D014162:Transfection; D001616:beta-Galactosidase / Q000378:metabolism
D004676:Myelin Basic Protein; D003850:Deoxyribonuclease I; D001616:beta-Galactosidase