Takanaga H, Mukai H, Shimakawa M, Konishi H, Kikkawa U, Koizumi T, Ono Y
Department of Biology, Faculty of Science, Kobe University, Japan.
FEBS Lett. 1995 Jul 17;368(2):276-8. doi: 10.1016/0014-5793(95)00665-v.
Promoter activity of protein kinase C (PKC) gamma gene was analysed by chloramphenicol acetyltransferase (CAT) assay using extracts from the cells transfected with various fusion constructs containing the 5'-flanking region of the mouse PKC gamma gene and CAT gene. Transient expression experiments in PC12 cells revealed that the upstream region of 87 bp from the transcriptional initiation site was sufficient for promoter activity. The region containing nucleotides 87 upstream from the transcriptional initiation site was shown to silence CAT activity in Balb/c3T3 cells, in which mRNA of PKC gamma was not detected, suggesting that this region might contain a transcriptional regulatory element for the cell type-specific expression of the PKC gamma gene.
利用氯霉素乙酰转移酶（CAT）分析方法，通过用含有小鼠蛋白激酶C（PKC）γ基因5'侧翼区和CAT基因的各种融合构建体转染细胞所获得的提取物，来分析PKCγ基因的启动子活性。在PC12细胞中的瞬时表达实验表明，转录起始位点上游87 bp的区域足以产生启动子活性。转录起始位点上游87个核苷酸的区域在Balb/c3T3细胞中显示会使CAT活性沉默，在该细胞中未检测到PKCγ的mRNA，这表明该区域可能含有PKCγ基因细胞类型特异性表达的转录调控元件。
