Peterson D S, Miller L H, Wellems T E
Laboratory of Parasitic Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892, USA.
Proc Natl Acad Sci U S A. 1995 Jul 18;92(15):7100-4. doi: 10.1073/pnas.92.15.7100.
Open reading frames in the Plasmodium falciparum genome encode domains homologous to the adhesive domains of the P. falciparum EBA-175 erythrocyte-binding protein (eba-175 gene product) and those of the Plasmodium vivax and Plasmodium knowlesi Duffy antigen-binding proteins. These domains are referred to as Duffy binding-like (DBL), after the receptor that determines P. vivax invasion of Duffy blood group-positive human erythrocytes. Using oligonucleotide primers derived from short regions of conserved sequence, we have developed a reverse transcription-PCR method that amplifies sequences encoding the DBL domains of expressed genes. Products of these reverse transcription-PCR amplifications include sequences of single-copy genes (including eba-175) and variably transcribed genes that cross-hybridize to multiple regions of the genome. Restriction patterns of the multicopy genes show a high degree of polymorphism among different parasite lines, whereas single-copy genes are generally conserved. Characterization of the single-copy genes has identified a gene (ebl-1) that is related to eba-175 and is likely to be involved in erythrocyte invasion.
恶性疟原虫基因组中的开放阅读框编码的结构域,与恶性疟原虫EBA - 175红细胞结合蛋白(eba - 175基因产物)以及间日疟原虫和诺氏疟原虫达菲抗原结合蛋白的黏附结构域同源。这些结构域被称为达菲结合样(DBL)结构域,得名于决定间日疟原虫侵入达菲血型阳性人类红细胞的受体。利用从保守序列短区域衍生而来的寡核苷酸引物,我们开发了一种逆转录 - PCR方法,该方法可扩增编码表达基因DBL结构域的序列。这些逆转录 - PCR扩增产物包括单拷贝基因(包括eba - 175)的序列以及与基因组多个区域交叉杂交的可变转录基因的序列。多拷贝基因的限制性酶切图谱在不同寄生虫株之间显示出高度多态性,而单拷贝基因通常是保守的。对单拷贝基因的表征鉴定出一个与eba - 175相关的基因(ebl - 1),它可能参与红细胞入侵。