Maingon R, Gerke R, Rodriguez M, Urbina J, Hoenicka J, Negri S, Aguirre T, Nehlin J, Knapp T, Crampton J
Departamento de Biologia Celular, Universidad Simon Bolivar, Caracas.
Eur J Biochem. 1988 Jan 15;171(1-2):285-91. doi: 10.1111/j.1432-1033.1988.tb13788.x.
The organization of the alpha- and beta-tubulin genes in the genome of Trypanosoma cruzi have been analysed by Southern blotting using tubulin probes derived from Trypanosoma brucei. The tubulin array appears to be more complex in this organism than in other members of the same family. Some tubulin genes are tightly clustered in an alternating (alpha-beta)n array with a basic repeat unit length of 4.3 kb. However, other pairs of alternating alpha- and beta-tubulin sequences appear to be physically separated from the basic group. This finding indicates that the tubulin gene cluster present in T. cruzi is less perfectly conserved than in T. brucei. T. (Herpetosoma) rangeli is similar to T. (Schizotrypanum) cruzi in its tubulin gene organization whereas most of these genes are tandemly clustered in the genome of T. (Trypanozoon) evansi, with a basic repeat unit length of 3.6 kb as previously described for T. (Trypanozoon) brucei. Two overlapping recombinant clones containing T. cruzi tubulin sequences have been isolated from a genomic cosmid library of T. cruzi epimastigotes using the T. brucei tubulin probes. Partial sequencing of the T. cruzi beta-tubulin gene has confirmed its identity and shows more than 70% homology with the sea urchin, chicken and T. b. rhodesiense beta-tubulin reported gene sequences. Analysis of tubulin gene organization through the parasite life cycle does not show evidence of major rearrangements within the repeat unit. Several T. cruzi strains and cloned lines whilst sharing the 4.3-kb tubulin repeat unit, exhibited very variable tubulin gene organization with tubulin probes. These striking differences in the organization of this structural gene among T. cruzi strains and cloned lines suggest that the heterogeneity previously reported in parasite populations may be related to a very dynamic, diploid genome.
利用源自布氏锥虫的微管蛋白探针,通过Southern杂交技术分析了克氏锥虫基因组中α-和β-微管蛋白基因的组织情况。该生物体中的微管蛋白阵列似乎比同一家族的其他成员更为复杂。一些微管蛋白基因紧密聚集在一个交替的(α-β)n阵列中,基本重复单元长度为4.3 kb。然而,其他成对的交替α-和β-微管蛋白序列似乎在物理上与基本组分离。这一发现表明,克氏锥虫中存在的微管蛋白基因簇不如布氏锥虫中那样保守。兰氏锥虫(赫佩托体锥虫)在微管蛋白基因组织方面与克氏锥虫(裂体锥虫)相似,而这些基因中的大多数在伊氏锥虫(锥虫亚属)的基因组中串联聚集,基本重复单元长度为3.6 kb,这与先前描述的布氏锥虫(锥虫亚属)相同。使用布氏锥虫微管蛋白探针,从克氏锥虫上鞭毛体的基因组黏粒文库中分离出了两个包含克氏锥虫微管蛋白序列的重叠重组克隆。克氏锥虫β-微管蛋白基因的部分测序证实了其身份,并显示与海胆、鸡以及布氏罗得西亚锥虫的β-微管蛋白已报道基因序列具有70%以上的同源性。通过寄生虫生命周期对微管蛋白基因组织的分析未显示重复单元内有重大重排的证据。几个克氏锥虫菌株和克隆系虽然共享4.3 kb的微管蛋白重复单元,但在用微管蛋白探针检测时,显示出非常多变的微管蛋白基因组织。克氏锥虫菌株和克隆系之间这种结构基因组织的显著差异表明,先前在寄生虫群体中报道的异质性可能与一个非常动态的二倍体基因组有关。