Ho L C, Armiugam A, Jeyaseelan K, Yap E H, Singh M
Department of Microbiology, Faculty of Medicine, National University of Singapore, Singapore.
Parasitology. 2000 Aug;121 ( Pt 2):135-44. doi: 10.1017/s0031182099006113.
The elongation factor-1 alpha (EF-1alpha) is a highly conserved ubiquitous protein that is involved in translation and is desirable for use in phylogenetic studies on Blastocystis, an enigmatic intestinal parasite with a contentious taxonomic position. In the present study, a PCR product (BEalpha) that codes for a major part of the coding region of the EF-lalpha protein was amplified. Genome walking experiments together with cloning were implemented to elucidate the 5' and 3' ends of the EF-1alpha gene of the human isolate, Blastocystis hominis C. The genomic organization and the potential transcription factor binding sites of the 5' end of B. hominis C EF-1alpha were identified. A comparative study on the deduced amino acid sequences of BEalpha of 13 Blastocystis isolates from various hosts was done to evaluate the phylogenetic relationship among the species. A phylogenetic reconstruction analysis with other eukaryotic EF-1alpha sequences was carried out to trace the phylogenetic position of Blastocystis among eukaryotic organisms.
延伸因子-1α(EF-1α)是一种高度保守的普遍存在的蛋白质,参与翻译过程,适用于对芽囊原虫进行系统发育研究。芽囊原虫是一种神秘的肠道寄生虫,其分类地位存在争议。在本研究中,扩增出了编码EF-1α蛋白编码区主要部分的PCR产物(BEα)。进行了基因组步移实验并结合克隆,以阐明人源分离株人芽囊原虫C的EF-1α基因的5'和3'末端。鉴定了人芽囊原虫C EF-1α 5'末端的基因组结构和潜在转录因子结合位点。对来自不同宿主的13株芽囊原虫分离株的BEα推导氨基酸序列进行了比较研究,以评估物种间的系统发育关系。利用其他真核生物EF-1α序列进行了系统发育重建分析,以追溯芽囊原虫在真核生物中的系统发育位置。