McGarvey M J, Sheybani E, Loche M P, Perrin L, Mach B
Proc Natl Acad Sci U S A. 1984 Jun;81(12):3690-4. doi: 10.1073/pnas.81.12.3690.
The key steps in the development of a malaria vaccine through gene cloning are the identification of the proteins involved in host protective immunity and the cloning, identification, and expression of the genes coding for these proteins. Recent data have indicated that certain proteins synthesized at the late schizont-merozoite stage of Plasmodium falciparum play a major role in malaria immunity. This paper reports the identification, in a cDNA library, of recombinant clones corresponding to genes expressed specifically during the late schizont-merozoite stage of P. falciparum development. The 132 cDNA clones thus identified out of 10,000 were found to correspond to only 12 different genes, probably representing most of the major schizont-merozoite specific genes. The stage-specific cDNAs can be efficiently expressed in Escherichia coli cells. The protein products of some of these clones are recognized by monoclonal antibodies specific for late schizont-merozoite proteins. We conclude that only a small set of genes is specifically induced in the schizont-merozoite stage and that the stage-specific cDNA clones we have isolated are very likely to include the genes coding for the immunologically relevant proteins of P. falciparum.
通过基因克隆研发疟疾疫苗的关键步骤包括确定参与宿主保护性免疫的蛋白质，以及对编码这些蛋白质的基因进行克隆、鉴定和表达。最近的数据表明，恶性疟原虫裂殖体-子孢子后期合成的某些蛋白质在疟疾免疫中起主要作用。本文报道了在一个cDNA文库中鉴定出与恶性疟原虫发育后期裂殖体-子孢子阶段特异性表达的基因相对应的重组克隆。在10000个克隆中鉴定出的这132个cDNA克隆被发现仅对应12个不同的基因，可能代表了大多数主要的裂殖体-子孢子特异性基因。这些阶段特异性cDNA能在大肠杆菌细胞中高效表达。其中一些克隆的蛋白质产物可被针对裂殖体-子孢子后期蛋白质的单克隆抗体识别。我们得出结论，在裂殖体-子孢子阶段仅诱导了一小部分基因，并且我们分离出的阶段特异性cDNA克隆很可能包含编码恶性疟原虫免疫相关蛋白质的基因。
