Tannich E, Scholze H, Nickel R, Horstmann R D
Bernhard Nocht Institute for Tropical Medicine, Hamburg, Federal Republic of Germany.
J Biol Chem. 1991 Mar 15;266(8):4798-803.
A cDNA clone derived from the gene encoding a cysteine proteinase of pathogenic Entamoeba histolytica was isolated using an antiserum to the purified enzyme. This clone was used to identify the homologous clone in a cDNA library from nonpathogenic E. histolytica. Sequence analysis and comparison of the predicted amino acid sequences revealed a sequence divergence of 16%. Southern blot analyses indicated that (i) pathogenic isolates may contain more genes coding for these or related enzymes than nonpathogenic isolates, (ii) the structure and organization of these genes are conserved within each group of amoebae, and (iii) none of the genes is found in both pathogenic and nonpathogenic E. histolytica, underlining the notion that the two groups are genetically distinct. Northern blot analyses suggested that the cysteine proteinase is expressed by pathogenic isolates in substantially higher amounts than by nonpathogenic isolates. Overexpression of this enzyme may be an important factor in the pathogenicity of E. histolytica.
利用针对纯化酶的抗血清,分离出了一个来自编码致病性溶组织内阿米巴半胱氨酸蛋白酶基因的cDNA克隆。该克隆用于鉴定非致病性溶组织内阿米巴cDNA文库中的同源克隆。对预测氨基酸序列的序列分析和比较显示,序列差异为16%。Southern印迹分析表明:(i)致病性分离株可能比非致病性分离株含有更多编码这些或相关酶的基因;(ii)这些基因的结构和组织在每组阿米巴中是保守的;(iii)在致病性和非致病性溶组织内阿米巴中均未发现相同的基因,这突出了两组在遗传上不同的观点。Northern印迹分析表明,致病性分离株表达的半胱氨酸蛋白酶的量远高于非致病性分离株。这种酶的过度表达可能是溶组织内阿米巴致病性的一个重要因素。