Malaria vaccine development of Plasmodium vivax, by using ookinete surface proteins as vaccine antigens

使用动动表面蛋白作为疫苗抗原开发间日疟原虫疟疾疫苗

基本信息

批准号：
09670261
负责人：
TSUBOI Takafumi
金额：
$ 2.3万
依托单位：
Ehime University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1998
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09670261/
关键词：
malaria vaccine Plasmodium vivax transmission-blocking gene cloning オ-キネート

项目摘要

In many malarious regions outside of Africa, development of transmission-blocking vaccine will require activity against both Plasimodiun falciparum and P.vivax. Work on P.vivax transmission-blocking vaccines have been hampered by the inability to clone the vaccine candidate genes from this parasite. To scarch for genes encoding the ookinete surface proteins from P.vivax, the gene sequences of tile eight known proteins in P25 subfamily (Pfs25, Pgs25, Pys25, Pbs25) and in P21/28 subfamily (Pfs28, Pgs28, Pys2l, Pbs2l) were aligned. Regions of highest identity were used to design degenerate PCR oligonucleotides. P.vivax Sail genomic DNA and genomic and splinkerette DNA libraries were used as PCR templates. Analysis of tile deduced amino acid sequence of Pvs28 revealed a secretory signal sequence, four EGF-like domains, six copies of the heptad amino acid repeat (GSGGE/D) and then a short hydrophobic region. Pvs28 is the presumed homologue of P21/28 subfamily member because the fourth EGF-like domain has four rather than six cysteines. Analysis of the deduced amino acid sequence of Pvs25 revealed a similar structure to Pvs28. Tile presence of six rather than four cysteines in tile fourth EGF-like domain suggested that Pvs25 is the homologue of P25 subfamily member. Accordingly, we have successfully cloned novel ookinete surface protein genes, Pvs28 and Pvs25, as the transmission-blocking vaccille candidate antigens, from P.vivax.

在非洲以外的许多疟疾流行地区，开发阻断传播的疫苗需要同时针对恶性疟原虫和间日疟原虫。由于无法从这种寄生虫中克隆疫苗候选基因，开发阻断间日疟原虫传播的疫苗的工作受到了阻碍。为了寻找编码间日疟原虫动动表面蛋白的基因，对 P25 亚家族 (Pfs25、Pgs25、Pys25、Pbs25) 和 P21/28 亚家族 (Pfs28、Pgs28、Pys2l、Pbs2l) 中的 8 个已知蛋白的基因序列进行了分析。对齐。最高同一性的区域被用来设计简并 PCR 寡核苷酸。 P.vivax Sail 基因组 DNA 以及基因组和 splinkerette DNA 文库用作 PCR 模板。对 Pvs28 推导的氨基酸序列的分析揭示了一个分泌信号序列、四个 EGF 样结构域、六个七肽氨基酸重复拷贝 (GSGGE/D) 和一个短疏水区域。 Pvs28 被认为是 P21/28 亚家族成员的同源物，因为第四个 EGF 样结构域具有四个而不是六个半胱氨酸。对 Pvs25 推导的氨基酸序列的分析揭示了与 Pvs28 相似的结构。第四个 EGF 样结构域中六个而不是四个半胱氨酸的存在表明 Pvs25 是 P25 亚家族成员的同源物。因此，我们成功地从间日疟原虫中克隆了新的动动表面蛋白基因Pvs28和Pvs25，作为阻断传播的疫苗候选抗原。