Shani M, Nudel U, Zevin-Sonkin D, Zakut R, Givol D, Katcoff D, Carmon Y, Reiter J, Frischauf A M, Yaffe D
Nucleic Acids Res. 1981 Feb 11;9(3):579-89. doi: 10.1093/nar/9.3.579.
Plasmids p749, p106, and p150 contain cDNA inserts complementary to rat skeletal muscle actin mRNA. Nucleotide sequence analysis indicates the following sequence relationships: p749 specifies codons 171 to 360; p150 specifies codons 357 to 374 together with 120 nucleotides of the 3'-non-translated region; p106 specifies the last actin amino acid codon, the termination codon and the entire 3' non-translated region. Plasmid p749 hybridized with RNA extracted from rat skeletal muscle, cardiac muscle, smooth (stomach) muscle, and from brain. It also hybridizes well with RNA extracted from skeletal muscle and brain of dog and chick. Plasmid p106 hybridized specifically with rat striated muscles (skeletal and cardiac muscle) mRNA but not with mRNA from rat stomach and from rat brain. It also hybridized to RNA extracted from skeletal muscle of rabbit and dog but not from chick. Thermal stability of the hybrids and sensitivity to S1 digestion also indicated substantial divergence between the 3' untranslated end of rat and dog skeletal muscle actins. The investigation shows that the coding regions of actin genes are highly conserved, whereas the 3' non-coding regions diverged considerably during evolution. Probes constructed from the 3' non-coding regions of actin mRNAs can be used to identify the various actin mRNA and actin genes.
质粒p749、p106和p150含有与大鼠骨骼肌肌动蛋白mRNA互补的cDNA插入片段。核苷酸序列分析表明了以下序列关系:p749编码第171至360个密码子;p150编码第357至374个密码子以及3'非翻译区的120个核苷酸;p106编码肌动蛋白的最后一个氨基酸密码子、终止密码子以及整个3'非翻译区。质粒p749与从大鼠骨骼肌、心肌、平滑肌(胃)以及脑提取的RNA杂交。它也能与从狗和鸡的骨骼肌和脑提取的RNA很好地杂交。质粒p106与大鼠横纹肌(骨骼肌和心肌)的mRNA特异性杂交,但不与大鼠胃和脑的mRNA杂交。它也能与从兔和狗的骨骼肌提取的RNA杂交,但不能与鸡的杂交。杂交体的热稳定性和对S1消化的敏感性也表明大鼠和狗骨骼肌肌动蛋白3'非翻译末端之间存在显著差异。研究表明,肌动蛋白基因的编码区高度保守,而3'非编码区在进化过程中发生了相当大的分化。由肌动蛋白mRNA的3'非编码区构建的探针可用于鉴定各种肌动蛋白mRNA和肌动蛋白基因。