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Genome-wide screening of the novel malaria transmission-blocking vaccine candidates

对新型疟疾传播阻断候选疫苗进行全基因组筛选

基本信息

批准号：
16390125
负责人：
TSUBOI Takafumi
金额：
$ 9.6万
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：
2004
资助国家：
日本
起止时间：
2004 至 2005
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-16390125/
关键词：
malaria vaccine genome-wide recombinant protein synthesis transmission-blocking International scientist exchange Thailand

项目摘要

Malaria remains one of the leading causes of both morbidity and mortality of humans residing in the tropical countries. The evidence of increasing resistance of the Plasmodium falciparum parasite to chemotherapeutic agents, such as chloroquine, highlights the critical need for an effective vaccine. However, the effective malaria vaccine has never been achieved to date. Since the genomic sequence of P.falciparum together with stage-specific proteome data was completed in October 2002, we have now free access to the genome data to search for novel vaccine candidates. However, one of the bottlenecks for the vaccine development is the high AT content in exons of P.falciparum, which will considerably inhibit the recombinant protein expression using conventional methods. Our strategy is to express recombinant proteins for the characterization of each protein based on the genome sequences of P.falciparum without using synthetic gene. Transmission-blocking vaccines (TBVs) prevent the transmiss … More ion of malaria by inducing antibodies against antigens specifically expressed on the sexual stage parasites. Since well-characterized TBV candidates are only four (Pfs25, Pfs28, Pfs48/45, and Pfs230), it would be necessary to prepare novel TBV candidates as many as possible for a successful TBV development. In order to identify the novel TBV candidates, we searched a combined dataset from genome and transcriptome databases and we selected 192 genes, which are expected to be expressed only in gametocyte stage of P.falciparum. These genes were cloned into plasmids and templates were prepared for transcription through PCR-based procedures, followed by high throughput recombinant protein synthesis by wheat germ cell-free system. Using this approach, we succeeded in obtaining 120 recombinant proteins. After the screening of these recombinant proteins to identify novel TBV candidates with anti-gametocyte monoclonal antibodies, we have identified a novel antigen expressed on the surface of gametocyte stage. Less

疟疾仍然是热带国家居民发病和死亡的主要原因之一。恶性疟原虫寄生虫对氯喹等化疗剂的抗药性不断增加的证据突出表明，迫切需要一种有效的疫苗。然而，迄今为止从未研制出有效的疟疾疫苗。由于恶性疟原虫的基因组序列和阶段特异性蛋白质组数据于2002年10月完成，我们现在可以免费获得基因组数据来寻找新的候选疫苗。然而，恶性疟原虫外显子中AT含量高，用常规方法表达重组蛋白会受到很大的抑制，这是恶性疟原虫疫苗研制的瓶颈之一。我们的策略是在不使用人工合成基因的情况下，基于恶性疟原虫基因组序列表达重组蛋白，以鉴定每种蛋白。传播阻断疫苗（TBVs）可预防传播 ...更多信息通过诱导针对在有性阶段寄生虫上特异表达的抗原的抗体来治疗疟疾。由于具有良好特征的TBV候选物只有四种（Pfs 25、Pfs 28、Pfs 48/45和Pfs 230），因此有必要为成功的TBV开发准备尽可能多的新TBV候选物。为了确定新的TBV候选人，我们从基因组和转录组数据库中搜索了一个组合数据集，我们选择了192个基因，预计仅在恶性疟原虫配子体阶段表达。将这些基因克隆到质粒中，并通过基于PCR的程序制备用于转录的模板，然后通过麦胚无细胞系统进行高通量重组蛋白合成。利用这种方法，我们成功地获得了120个重组蛋白。在用抗配子体单克隆抗体筛选这些重组蛋白以鉴定新的TBV候选物之后，我们鉴定了在配子体阶段表面表达的新抗原。少