Kretsovali A, Müller M M, Weber F, Marcaud L, Farache G, Schreiber E, Schaffner W, Scherrer K
Institute Jacques Monod, Paris, France.
Gene. 1987;58(2-3):167-75. doi: 10.1016/0378-1119(87)90373-8.
We have detected a transcriptional enhancer sequence downstream from the adult beta-globin (beta A-globin) genes of chicken and duck. DNA segments from the beta-globin coding and flanking sequences were cloned into expression vectors containing the SV40 promoter linked to either the T antigen gene or the cat gene. The expression of these genes was measured in a chicken erythroid cell line transfected with the recombinant plasmids. We found that segments located about 400 bp downstream from the poly(A) site of both the chicken and duck beta A-globin genes (and about 1.5 kb upstream from the embryonic epsilon-globin gene) stimulate transcription of the test genes about five-fold. In chicken essentially the same segment was also found by others to act as an erythroid cell-specific enhancer [Hesse et al., Proc. Natl. Acad. Sci. USA 83 (1986) 4312-4316; Choi and Engel, Nature 323 (1986) 731-734]. The sequence containing this enhancer is conserved in evolution. A high degree of homology, reaching 84% in a segment 180 bp in length, was found between chicken and duck despite an evolutionary divergence of 70 myr.
我们在鸡和鸭的成年β-珠蛋白(βA-珠蛋白)基因下游检测到一个转录增强子序列。将β-珠蛋白编码及侧翼序列的DNA片段克隆到含有与T抗原基因或氯霉素乙酰转移酶(cat)基因相连的SV40启动子的表达载体中。在转染了重组质粒的鸡红细胞系中检测这些基因的表达。我们发现,位于鸡和鸭βA-珠蛋白基因poly(A)位点下游约400 bp处(以及胚胎ε-珠蛋白基因上游约1.5 kb处)的片段可使测试基因的转录增强约5倍。在鸡中,其他人也发现基本相同的片段可作为红细胞特异性增强子[黑塞等人,《美国国家科学院院刊》83 (1986) 4312 - 4316;崔和恩格尔,《自然》323 (1986) 731 - 734]。含有该增强子的序列在进化过程中是保守的。尽管鸡和鸭在进化上有7000万年的分歧,但在长度为180 bp的片段中发现它们具有高达84%的高度同源性。