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首页> 外文期刊>Molecular and Cellular Biology >Cloning and characterization of the gene for beta-tubulin from a benomyl-resistant mutant of Neurospora crassa and its use as a dominant selectable marker.
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Cloning and characterization of the gene for beta-tubulin from a benomyl-resistant mutant of Neurospora crassa and its use as a dominant selectable marker.

机译：神经孢子菌苯致密突变体的粘附性和表征β-微管蛋白及其用作优势选择标记。

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摘要

We cloned the beta-tubulin gene of Neurospora crassa from a benomyl-resistant strain and determined its nucleotide sequence. The gene encodes a 447-residue protein which shows strong homology to other beta-tubulins. The coding region is interrupted by six introns, five of which are within the region coding for the first 54 amino acids of the protein. Intron position comparisons between the N. crassa gene and other fungal beta-tubulin genes reveal considerable positional conservation. The mutation responsible for benomyl resistance was determined; it caused a phenylalanine-to-tyrosine change at position 167. Codon usage in the beta-tubulin gene is biased, as has been observed for other abundantly expressed N. crassa genes such as am and the H3 and H4 histone genes. This bias results in pyrimidines in the third positions of 96% of the codons in codon families in which there is a choice between purines and pyrimidines in this position. Bias is also evident by the absence of 19 of the 61 sense codons. We demonstrated that benomyl resistance is due to the cloned beta-tubulin gene of strain Bml511(r)a and that this gene can be used as a dominant selectable marker in N. crassa transformation.
机译：我们克隆了来自苯甲瘤菌株的神经孢子塔的β-微管蛋白基因并确定其核苷酸序列。该基因编码了447-残基蛋白，其显示对其他β-管蛋白的强烈同源性。编码区域被六个内含子中断，其中5个内部在编码蛋白质的前54个氨基酸的区域内。 N.Crassa基因与其他真菌β-微管蛋白基因之间的内含子位置比较揭示了相当大的位置保护。测定负责苯甲基抗性的突变;它导致在第167位的苯丙氨酸 - 酪氨酸变化。β-微管蛋白基因的密码子使用偏置，如已经观察到的其他大规模表达的N.Crassa基因如AM和H3和H4组蛋白基因。这种偏差导致嘧啶在密码子系列中96％的密码子中的第三位，其中在该位置中存在嘌呤和嘧啶之间的选择。在没有191个感测密码子中，偏差也是显而易见的。我们证明，苯甲基抗性是由于菌株BML511（R）A的克隆β-微管蛋白基因，并且该基因可以用作N.Crassa转化中的显性选择标记。

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