Grichnik J M, Bergsma D J, Schwartz R J
Nucleic Acids Res. 1986 Feb 25;14(4):1683-701. doi: 10.1093/nar/14.4.1683.
alpha-skeletal actin message levels have been shown to be tightly regulated in chicken primary myoblast cultures. To test for gene elements required for muscle cell specific expression, DNA sequences containing the 5'-flanking regions of the chicken alpha-skeletal actin, beta-cytoplasmic actin, and the histone H2b genes were linked to the coding sequences of the chloramphenicol acetyltransferase gene and transfected into myogenic and non-myogenic cells. In contrast to beta-actin CAT hybrids, the alpha-skeletal actin CAT constructions displayed restricted CAT expression in transfected non-myogenic cells. We showed that a 411 nucleotide fragment flanking the 5' end of of the alpha-skeletal actin gene was responsible for a 9-15 fold increase in CAT enzymatic activity during myoblast fusion, versus only a transient 2 fold rise for the beta-actin and histone flanking sequences. These results indicate that DNA sequences within 411 bp of the 5' terminus of the alpha-skeletal actin gene influenced its cell type and stage specific expression.
已证明α-骨骼肌动蛋白的信息水平在鸡原代成肌细胞培养物中受到严格调控。为了测试肌肉细胞特异性表达所需的基因元件，将包含鸡α-骨骼肌动蛋白、β-细胞质动蛋白和组蛋白H2b基因5'侧翼区域的DNA序列与氯霉素乙酰转移酶基因的编码序列相连，并转染到成肌细胞和非成肌细胞中。与β-肌动蛋白氯霉素乙酰转移酶杂种不同，α-骨骼肌动蛋白氯霉素乙酰转移酶构建体在转染的非成肌细胞中显示出有限的氯霉素乙酰转移酶表达。我们表明，α-骨骼肌动蛋白基因5'端侧翼的一个411核苷酸片段在成肌细胞融合过程中导致氯霉素乙酰转移酶酶活性增加9至15倍，而β-肌动蛋白和组蛋白侧翼序列仅短暂增加2倍。这些结果表明，α-骨骼肌动蛋白基因5'末端411 bp内的DNA序列影响其细胞类型和阶段特异性表达。
