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担子菌裂褶菌中β-微管蛋白编码基因的克隆、测序及表达

Cloning, sequence and expression of a beta-tubulin-encoding gene in the homobasidiomycete Schizophyllum commune.

作者信息

Russo P, Juuti J T, Raudaskoski M

机构信息

Department of Botany, University of Helsinki, Finland.

出版信息

Gene. 1992 Oct 1;119(2):175-82. doi: 10.1016/0378-1119(92)90269-u.

Abstract

The beta-tubulin (beta Tub)-encoding gene (tub-2) of Schizophyllum commune is the first tubulin gene isolated, cloned and sequenced from higher filamentous fungi (homobasidiomycetes). The S. commune tub-2 gene is organized into nine exons and eight introns. The introns vary from 48 to 107 nt in length, and are distributed throughout the gene. The tub-2 exons code for a protein of 445 amino acids (aa), which shows great homology with beta Tubs of filamentous ascomycetes, plants, and animals, but less homology with yeasts. The codon usage of tub-2 from S. commune is biased, as it is in most beta Tub-encoding genes of filamentous fungi. The S. commune beta Tub shows a conserved aa sequence in the C-terminal domain, which is suggested to interact with microtubule-associated proteins in animals. In contrast, the S. commune beta Tub deviates from most known beta Tubs by having a Cys165 residue, which might be significant for the insensitivity of S. commune haploid strains to the antimicrotubule drug, benomyl. In tub-2 of different haploid strains, sequence polymorphisms occur in the 5' and 3' flanking regions. The expression of tub-2 is high in young mycelium, which has a high number of extending apical cells, but decreases with the aging of the mycelium. No significant difference in the hybridization signal intensity for the tub-2 transcripts was recorded either during intercellular nuclear migration at early mating, or in mycelia with a mutation in the B mating-type gene.(ABSTRACT TRUNCATED AT 250 WORDS)

摘要

裂褶菌的β-微管蛋白(β Tub)编码基因(tub-2)是从高等丝状真菌(同担子菌)中分离、克隆和测序的首个微管蛋白基因。裂褶菌tub-2基因由9个外显子和8个内含子组成。内含子长度从48到107个核苷酸不等,分布于整个基因中。tub-2外显子编码一个由445个氨基酸组成的蛋白质,该蛋白质与丝状子囊菌、植物和动物的β Tub具有高度同源性,但与酵母的同源性较低。裂褶菌tub-2的密码子使用存在偏向性,大多数丝状真菌的β Tub编码基因也是如此。裂褶菌β Tub在C末端结构域显示出保守的氨基酸序列,提示其与动物中的微管相关蛋白相互作用。相比之下,裂褶菌β Tub与大多数已知的β Tub不同,它有一个Cys165残基,这可能是裂褶菌单倍体菌株对抗微管药物苯菌灵不敏感的原因。在不同单倍体菌株的tub-2中,5'和3'侧翼区域存在序列多态性。tub-2在具有大量延伸顶端细胞的幼菌丝体中表达较高,但随着菌丝体老化而降低。在早期交配时的细胞间核迁移过程中,或者在B交配型基因突变的菌丝体中,tub-2转录本的杂交信号强度均未记录到显著差异。(摘要截断于250字)

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