Expression of Plasmodium falciparum surface antigens in Escherichia coli.
Expression of Plasmodium falciparum surface antigens in Escherichia coli.
复制标题
恶性疟原虫表面抗原在大肠杆菌中的表达。
DOI：
10.1073/pnas.82.8.2518
复制
发表时间：
1985
影响因子：
11.1
通讯作者：
Reese,RT
中科院分区：
文献类型：
--
作者：
Ardeshir,F;Flint,JE;Reese,RT
人类疟原虫恶性疟原虫的无性血液阶段产生许多抗原，其中只有一些是重要的保护性免疫。大多数假定的保护性抗原被认为在无性生殖周期的晚期阶段的裂殖子和裂殖子中表达。为了克隆和表征重要寄生虫抗原的基因，我们使用晚期恶性疟原虫mRNA构建插入大肠杆菌表达载体pUC 8中的cDNA序列文库。从表达文库中筛选出9000个克隆，用抗恶性疟原虫的Aotus猴血清进行原位免疫筛选，鉴定出95个表达疟原虫抗原的克隆。用来自49个与Aotus血清反应的细菌克隆的裂解物免疫小鼠，并通过间接免疫荧光、免疫沉淀和免疫印迹测定来测试小鼠血清与寄生虫抗原的反应性。通过这些测定鉴定了几种不同的恶性疟原虫抗原。细胞外裂殖子的间接免疫荧光研究表明，这些抗原中的三个似乎位于裂殖子表面。因此，我们已经确定了三种不同的恶性疟原虫抗原，可能是重要的保护性免疫的cDNA克隆。
The asexual blood stages of the human malarial parasite Plasmodium falciparum produce many antigens, only some of which are important for protective immunity. Most of the putative protective antigens are believed to be expressed in schizonts and merozoites, the late stages of the asexual cycle. With the aim of cloning and characterizing genes for important parasite antigens, we used late-stage P. falciparum mRNA to construct a library of cDNA sequences inserted in the Escherichia coli expression vector pUC8. Nine thousand clones from the expression library were immunologically screened in situ with serum from Aotus monkeys immune to P. falciparum, and 95 clones expressing parasite antigens were identified. Mice were immunized with lysates from 49 of the bacterial clones that reacted with Aotus sera, and the mouse sera were tested for their reactivity with parasite antigens by indirect immunofluorescence, immunoprecipitation, and immunoblotting assays. Several different P. falciparum antigens were identified by these assays. Indirect immunofluorescence studies of extracellular merozoites showed that three of these antigens appear to be located on the merozoite surface. Thus, we have identified cDNA clones to three different P. falciparum antigens that may be important in protective immunity.