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根据GenBank上已发表的其他真菌α–微管蛋白基因序列中的保守区域设计简并引物,扩增出银耳α–微管蛋白基因的cDNA部分序列。分别利用SEFA-PCR方法和RACE技术,克隆了银耳α–微管蛋白基因的DNA和cDNA全长序列。银耳α–微管蛋白基因DNA全长1962bp,含有9个内含子,cDNA全长1347bp,编码448个氨基酸。生物信息学分析结果表明:银耳α–微管蛋白基因的cDNA序列与新型隐球酵母有80%相似性,编码的蛋白质属于Tubulin_FtsZ超家族中的α–微管蛋白亚家族,氨基酸序列中存在保守的GTP结合区域GGGTGSG,系统发育树显示银耳与新型隐球酵母距离最近。
According to the conserved regions of the other fungal α-tubulin genes published in GenBank, degenerate primers were designed to amplify the cDNA partial sequence of the α-tubulin gene. The full-length DNA and cDNA sequences of α-tubulin gene were cloned by SEFA-PCR and RACE respectively. The DNA of T. auricularα-tubulin gene is 1962bp in length and contains 9 introns. The cDNA is 1347bp in length and encodes 448 amino acids. The results of bioinformatics analysis showed that the cDNA sequence of the α-tubulin gene was 80% similar to that of Cryptococcus neoformans, and the encoded protein belonged to α-tubulin subfamily of Tubulin_FtsZ superfamily. The amino acid sequence of the protein was conserved Of the GTP binding region GGGTGSG, phylogenetic tree shows white fungus and Cryptococcus neoformans from the recent.