Analysis of the protein expression level of the erythrocyte binding proteins in Plasmodium falciparum field isolates.

恶性疟原虫野外分离株中红细胞结合蛋白的蛋白表达水平分析。

• 批准号: 15406015
• 负责人: TORII Motomi
• 金额: $ 8.58万
• 依托单位: Ehime University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-15406015/
• 关键词: infectious disease; malaria; Plasmodium; マラリア; 熱帯熱マラリア; 赤血球結合蛋白; 微生物; 侵入; 遺伝子; infectious disease; malaria; Plasmodium; マラリア; 熱帯熱マラリア; 赤血球結合蛋白; 微生物; 侵入; 遺伝子

Erythrocyte binding proteins are potent malaria vaccine candidates and the expression of these proteins in the field isolates has to be evaluated before vaccine development based upon these proteins. The objective of this project is to analyze mRNA and protein expression for the erythrocyte binding proteins, encoded by multigene family EBL, RBL, and Clag/RhopH1, which are involved in the erythrocyte invasion process. We collected P.falciparum field isolates in Thailand and cultured for short period and enriched mature schizonts. Using oligonucleotide targeting members of each multigene family, RT-PCR was performed to detect transcripts level. However, by preliminal analysis of the polymorphism of these genes, we found these gene family members are highly diversified and carefull design of the oligonucleotide is required to distinguish each member, but identify individual members. We generated a panel of antisera against each members and analyzed the expression levels of the each member. For Clag/RhopH1, the region showing diversified each other among members shows also polymorphic among isolates, indicated that it is required to generate antisera which distinguish each members, but react all type of the allele.

红细胞结合蛋白是有效的疟疾疫苗候选物，基于这些蛋白质的疫苗发育之前，必须评估这些蛋白在田间分离株中的表达。该项目的目的是分析红细胞结合蛋白的mRNA和蛋白质表达，该蛋白质由多基因家族EBL，RBL和锁骨/Rhoph1编码，这些蛋白与红细胞浸润过程有关。我们在泰国收集了菲尔卡鲁姆的田间分离株，并在短时间内进行了培养，并富含成熟的精神分裂症。使用每个多基因家族的寡核苷酸靶向成员，进行了RT-PCR来检测转录物水平。但是，通过对这些基因的多态性的前分析，我们发现这些基因家族成员是高度多样化的，并且需要对寡核苷酸的仔细设计来区分每个成员，但要识别单个成员。我们针对每个成员生成了一组抗血小板，并分析了每个成员的表达水平。对于作作thag/Rhoph1的区域，在成员之间互相多样化的区域也显示了分离株之间的多态性，表明需要产生区分每个成员的抗血清，但反应所有类型的等位基因。