Effect of bacterial association on the phenotype and genotype of an Entamoeba histolytica clonal population.

Invasion & metastasis Pub Date : 1997-01-01
L F De Menezes, M A Rodríguez, M A Vargas, L M Salgado, E Orozco
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Abstract

A several-times-cloned population of Entamoeba histolytica trophozoites (clone MAVIII) was cultured under axenic (MAVIIIax), monoxenic (MAVIIImx) and polyxenic (MAVIIIpx) conditions. Clones MAVIIIax and MAVIIImx presented similar virulence in vitro, but differed in their virulence in vivo, whereas MAVIIIpx trophozoites were neither virulent in vitro or in vivo. The MAVIII clones maintained their zymodeme and exhibited three unusual glucose phosphate isomerase bands, absent in other E. histolytica strains studied. Similar patterns were shown by the three MAVIII clones in the signature of a 482-bp DNA fragment from the M17 gene (which encodes for a variable immunodominant antigen), obtained by low stringency single specific primer PCR technique. However, MAVIII clones displayed genotypic variability in the patterns obtained by the random amplified polymorphic DNA technique using total DNA as template. Results suggest that monomorphism is kept in certain regions of the genome, mainly in those carrying protein encoding genes, but a high polymorphism is present in total DNA of cloned trophozoites cultured under different conditions, confirming the plasticity of the E. histolytica genome.

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细菌关联对溶组织内阿米巴克隆群体表型和基因型的影响。
将多次克隆的溶组织内阿米巴滋养体(克隆MAVIII)在单株(maviiax)、单株(MAVIIImx)和多株(MAVIIIpx)条件下进行培养。maviiax和MAVIIImx克隆在体外毒力相似,但在体内毒力不同,而MAVIIIpx滋养体在体外和体内均无毒力。MAVIII克隆保持了它们的酶原,并表现出三个不同寻常的葡萄糖磷酸异构酶带,这在其他研究的溶组织芽胞杆菌菌株中是不存在的。通过低强度单特异性引物PCR技术获得的M17基因(编码可变免疫优势抗原)的482 bp DNA片段的特征也显示出类似的模式。然而,MAVIII克隆在以总DNA为模板的随机扩增多态性DNA技术获得的模式中显示出基因型变异。结果表明,在基因组的某些区域,主要是携带蛋白质编码基因的区域,保持了单态性,但在不同条件下培养的克隆滋养体的总DNA中存在高度多态性,证实了溶组织芽孢杆菌基因组的可塑性。
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