Elliott J F, Albrecht G R, Gilladoga A, Handunnetti S M, Neequaye J, Lallinger G, Minjas J N, Howard R J
DNAX Research Institute, Palo Alto, CA 94304-1104.
Proc Natl Acad Sci U S A. 1990 Aug;87(16):6363-7. doi: 10.1073/pnas.87.16.6363.
Two genes encoding membrane antigens of Plasmodium falciparum were isolated by transient expression in mammalian cells and selection with human immune sera from African adults exposed to P. falciparum malaria. COS-7 cells were transfected with a plasmid expression library constructed from P. falciparum genomic DNA, and cells expressing reactive malaria antigens on their surface were enriched by adherence to antibody-coated dishes. One of the genes isolated is distinctive in that it does not contain repeat sequences typical of many malarial genes cloned by immunoscreening of bacterial expression libraries. The second gene apparently encodes a polymorphic version of the P. falciparum merozoite surface antigen Ag513, since the two sequences are identical in the 5' and 3' coding regions but diverge completely in the center. The COS-7 expression system provides an alternate means for cloning genes encoding malarial membrane antigens by using those antibodies in complex immune sera that bind membrane-associated, nondenatured molecules.
通过在哺乳动物细胞中瞬时表达,并利用来自接触恶性疟原虫疟疾的非洲成年人的人免疫血清进行筛选,分离出了两个编码恶性疟原虫膜抗原的基因。用从恶性疟原虫基因组DNA构建的质粒表达文库转染COS-7细胞,通过粘附到抗体包被的培养皿上富集在其表面表达反应性疟疾抗原的细胞。分离出的其中一个基因很独特,因为它不包含通过对细菌表达文库进行免疫筛选克隆的许多疟疾基因典型的重复序列。第二个基因显然编码恶性疟原虫裂殖子表面抗原Ag513的多态形式,因为这两个序列在5'和3'编码区域相同,但在中间完全不同。COS-7表达系统提供了一种替代方法,通过使用复杂免疫血清中那些结合膜相关、未变性分子的抗体来克隆编码疟疾膜抗原的基因。