Boardman M, Cross G S, Jones E A, Woodland H R
Department of Biological Sciences, University of Warwick, Coventry, England.
Eur J Biochem. 1992 Sep 1;208(2):241-9. doi: 10.1111/j.1432-1033.1992.tb17179.x.
We have isolated a genomic clone, related in sequence to the skeletal-actin gene sub-family. It is expressed in the skeletal muscle of embryos from the neurula stage onwards and in tadpoles, but not in adults. The equivalent Xenopus laevis gene is expressed as a major transcript in adult muscle, as well as at earlier stages. The intron/exon structure is typical of vertebrate skeletal-actin genes, as is the possession of multiple copies of three serum-response elements in the promoter of this gene. The Xenopus actin and beta-globin genes were fused in their second introns. This construct, which contained 2.4 kb of upstream sequence, was injected into fertilized eggs at the two-cell stage. It showed the normal pattern of tissue-specific transcription. Thus all of the information necessary for appropriate expression of this actin gene in the embryo is contained in the region that extends from a point 2.4 kb upstream of transcription initiation to the centre of the second exon. A series of enhancer constructs were made in which upstream regions of the actin gene were placed upstream of a X. laevis beta-globin gene. The region immediately adjacent to the promoter, containing the three serum-response elements, was able to drive muscle-specific expression, and there was also a general enhancement of transcription by regions further upstream.
我们分离出了一个基因组克隆,其序列与骨骼肌肌动蛋白基因亚家族相关。它在神经胚阶段及之后的胚胎骨骼肌中表达,也在蝌蚪中表达,但在成体中不表达。非洲爪蟾的同源基因在成体肌肉以及更早阶段均作为主要转录本表达。该基因的内含子/外显子结构是脊椎动物骨骼肌肌动蛋白基因的典型结构,其启动子中拥有三个血清反应元件的多个拷贝也是如此。非洲爪蟾的肌动蛋白基因和β-珠蛋白基因在它们的第二个内含子处融合。这个包含2.4 kb上游序列的构建体在二细胞阶段被注射到受精卵中。它表现出正常的组织特异性转录模式。因此,该肌动蛋白基因在胚胎中正确表达所需的所有信息都包含在从转录起始点上游2.4 kb处到第二个外显子中心的区域内。构建了一系列增强子构建体,其中肌动蛋白基因的上游区域被置于非洲爪蟾β-珠蛋白基因的上游。紧邻启动子的区域,包含三个血清反应元件,能够驱动肌肉特异性表达,并且更上游的区域也普遍增强了转录。