Partington G A, Yarwood N J, Rutherford T R
EMBO J. 1984 Dec 1;3(12):2787-92. doi: 10.1002/j.1460-2075.1984.tb02210.x.
We have studied the expression of the human adult (beta) and foetal (gamma) globin genes by injecting a cosmid containing the complete -G gamma-A gamma-psi beta-delta-beta cluster into Xenopus oocytes. The transcripts from the gamma and beta genes were characterised with respect to their 5' termini by S1 nuclease mapping using probes which extend 5' to the cap site and 3' into the first exon. The only beta transcripts detectable in injected oocytes spanned from either -231 or -177 (cap site +1) into the first exon. The sensitivity of this transcription to inhibition by a low concentration of alpha-amanitin indicated that polymerase II was responsible. S1 analysis of gamma gene transcripts showed that the predominant transcript mapped from a site at -227, but a low level of transcription from the cap site was also detectable. Incubation of oocytes in saline solution containing sodium butyrate prior to injection resulted in a significant stimulation of transcription from the gamma gene cap site. In contrast, transcription from the beta gene upstream sites was only slightly increased. The possible role of sodium butyrate in promoting this effect is discussed in the context of its known property as a modifier of cellular chromatin.
我们通过将含有完整的-Gγ-Aγ-ψβ-δ-β基因簇的黏粒注射到非洲爪蟾卵母细胞中,研究了人类成人(β)和胎儿(γ)珠蛋白基因的表达。使用延伸至帽位点上游5'和进入第一个外显子3'的探针,通过S1核酸酶图谱分析,对γ和β基因的转录本5'末端进行了表征。在注射的卵母细胞中可检测到的唯一β转录本跨度为从-231或-177(帽位点+1)进入第一个外显子。这种转录对低浓度α-鹅膏蕈碱抑制的敏感性表明是聚合酶II负责。γ基因转录本的S1分析表明,主要转录本从-227位点定位,但也可检测到从帽位点的低水平转录。在注射前将卵母细胞在含有丁酸钠的盐溶液中孵育,导致γ基因帽位点的转录显著刺激。相比之下,β基因上游位点的转录仅略有增加。结合丁酸钠作为细胞染色质修饰剂的已知特性,讨论了丁酸钠促进这种效应的可能作用。