Silflow C D, Chisholm R L, Conner T W, Ranum L P
Mol Cell Biol. 1985 Sep;5(9):2389-98. doi: 10.1128/mcb.5.9.2389-2398.1985.
Full-length cDNA clones corresponding to the transcripts of the two alpha-tubulin genes in Chlamydomonas reinhardi were isolated. DNA sequence analysis of the cDNA clones and cloned gene fragments showed that each gene contains 1,356 base pairs of coding sequence, predicting alpha-tubulin products of 451 amino acids. Of the 27 nucleotide differences between the two genes, only two result in predicted amino acid differences between the two gene products. In the more divergent alpha 2 gene, a leucine replaces an arginine at amino acid 308, and a valine replaces a glycine at amino acid 366. The results predicted that two alpha-tubulin proteins with different net charges are produced as primary gene products. The predicted amino acid sequences are 86 and 70% homologous with alpha-tubulins from rat brain and Schizosaccharomyces pombe, respectively. Each gene had two intervening sequences, located at identical positions. Portions of an intervening sequence highly conserved between the two beta-tubulin genes are also found in the second intervening sequence of each of the alpha genes. These results, together with our earlier report of the beta-tubulin sequences in C. reinhardi, present a picture of the total complement of genetic information for tubulin in this organism.
分离出了与莱茵衣藻中两个α-微管蛋白基因转录本相对应的全长cDNA克隆。对cDNA克隆和克隆的基因片段进行DNA序列分析表明,每个基因包含1356个碱基对的编码序列,预测α-微管蛋白产物为451个氨基酸。在这两个基因之间的27个核苷酸差异中,只有两个导致两个基因产物之间预测的氨基酸差异。在差异更大的α2基因中,第308位氨基酸处的亮氨酸取代了精氨酸,第366位氨基酸处的缬氨酸取代了甘氨酸。结果预测,作为主要基因产物产生了两种带不同净电荷的α-微管蛋白。预测的氨基酸序列分别与大鼠脑和粟酒裂殖酵母的α-微管蛋白具有86%和70%的同源性。每个基因有两个间隔序列,位于相同位置。在每个α基因的第二个间隔序列中也发现了两个β-微管蛋白基因之间高度保守的间隔序列部分。这些结果,连同我们早期关于莱茵衣藻中β-微管蛋白序列的报告,展示了该生物体中微管蛋白遗传信息的完整图谱。