Detection of brain-specific gene expression in brain cells in primary culture: a novel promoter assay based on the use of a retrovirus vector.
Promoter activities of the brain-specific genes for glial fibrillary acidic protein (GFAP) and myelin basic protein (MBP) were investigated in brain cells in primary culture with the use of a novel retrovirus vector, pIP200. With this vector, promoter activity can be expressed in terms of beta-galactosidase activity. Differentiation of the primary brain cells to mature glial cells was not affected by treatment with the pIP200 virus vector. The 256-bp 5'-flanking region of the GFAP gene directed astrocyte-specific expression of lacZ. It was silent in fibroblasts, even in multiple copies. The 1.3-kb 5'-flanking region of the MBP gene exhibited strict tissue (oligodendrocyte) specificity under the present assay method but showed some leakiness when integrated into the chromosome in multiple copies. Promoter regions conferring cell type specificity in brain were effectively identified by the present method.
Ikenaka
K
K
Institute for Protein Research, Osaka University, Japan.
Nakahira
K
K
Nakajima
K
K
Fujimoto
I
I
Kagawa
T
T
Ogawa
M
M
Mikoshiba
K
K
D000818:Animals; D001483:Base Sequence; D001921:Brain / Q000502:physiology*; D002478:Cells, Cultured; D005786:Gene Expression Regulation / Q000502:physiology*; D005822:Genetic Vectors; D051379:Mice; D008813:Mice, Inbred ICR; D008969:Molecular Sequence Data; D009836:Oligodendroglia / Q000502:physiology; D010957:Plasmids / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics* / Q000502:physiology; D012190:Retroviridae / Q000235:genetics*