Use of high-density tiling microarrays to identify mutations globally and elucidate mechanisms of drug resistance in Plasmodium falciparum.
Use of high-density tiling microarrays to identify mutations globally and elucidate mechanisms of drug resistance in Plasmodium falciparum.
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DOI：
10.1186/gb-2009-10-2-r21
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发表时间：
2009-02-13
期刊：
影响因子：
12.3
通讯作者：
Winzeler EA
中科院分区：
文献类型：
--
作者：
Dharia NV;Sidhu AB;Cassera MB;Westenberger SJ;Bopp SE;Eastman RT;Plouffe D;Batalov S;Park DJ;Volkman SK;Wirth DF;Zhou Y;Fidock DA;Winzeler EA
使用平铺微阵列，恶性疟原虫寄生虫获得抗疟药磷咪霉素的耐药性的机制已被阐明。鉴定赋予病原体耐药性或其他表型变化的遗传变化可以帮助优化治疗策略，支持新治疗药物的开发，并提供有关基因可能功能的信息。阐明表型耐药性的机制也有助于确定在针对恶性疟原虫的高通量化学筛选活动中鉴定的未表征但有效的抗疟化合物的作用模式。在这里，我们表明，平铺微阵列可以从头检测到很大一部分的遗传变化，区分一个基因组从另一个。我们发现，我们检测到大多数单核苷酸多态性或小插入缺失事件和所有已知的拷贝数变异，区分三个实验室分离株使用方便的方法。我们使用这种方法来发现转染后选择过程中发生的突变。我们还阐明了寄生虫获得抗疟磷胺霉素，其目标是寄生虫类异戊二烯合成途径的耐药性的机制。我们的微阵列为基础的方法，使我们能够属性在体外衍生的磷咪霉素耐药的pfdxr基因，使寄生虫克服磷咪霉素介导的抑制类异戊二烯生物合成的拷贝数变异事件。我们发现，新出现的单核苷酸多态性可以很容易地检测到，疟原虫可以迅速获得基因扩增，在体外药物压力。通过一次过夜杂交全面确定恶性疟原虫遗传变异的能力为研究寄生虫进化和改善疟疾的治疗和控制创造了新的机会。
Using tiling microarrays, a mechanism by which Plasmodium falciparum parasites acquire resistance to the antimalarial fosmidomycin has been elucidated The identification of genetic changes that confer drug resistance or other phenotypic changes in pathogens can help optimize treatment strategies, support the development of new therapeutic agents, and provide information about the likely function of genes. Elucidating mechanisms of phenotypic drug resistance can also assist in identifying the mode of action of uncharacterized but potent antimalarial compounds identified in high-throughput chemical screening campaigns against Plasmodium falciparum. Here we show that tiling microarrays can detect de novo a large proportion of the genetic changes that differentiate one genome from another. We show that we detect most single nucleotide polymorphisms or small insertion deletion events and all known copy number variations that distinguish three laboratory isolates using readily accessible methods. We used the approach to discover mutations that occur during the selection process after transfection. We also elucidated a mechanism by which parasites acquire resistance to the antimalarial fosmidomycin, which targets the parasite isoprenoid synthesis pathway. Our microarray-based approach allowed us to attribute in vitro derived fosmidomycin resistance to a copy number variation event in the pfdxr gene, which enables the parasite to overcome fosmidomycin-mediated inhibition of isoprenoid biosynthesis. We show that newly emerged single nucleotide polymorphisms can readily be detected and that malaria parasites can rapidly acquire gene amplifications in response to in vitro drug pressure. The ability to define comprehensively genetic variability in P. falciparum with a single overnight hybridization creates new opportunities to study parasite evolution and improve the treatment and control of malaria.
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影响因子：
6.7
作者：
Ganesan K;Ponmee N;Jiang L;Fowble JW;White J;Kamchonwongpaisan S;Yuthavong Y;Wilairat P;Rathod PK
通讯作者：
Rathod PK
影响因子：
4.4
作者：
Jiang H;Yi M;Mu J;Zhang L;Ivens A;Klimczak LJ;Huyen Y;Stephens RM;Su XZ
通讯作者：
Su XZ
DOI：
10.1073/pnas.91.3.1143
发表时间：
1994-02-01
影响因子：
11.1
作者：
COWMAN, AF;GALATIS, D;THOMPSON, JK
通讯作者：
THOMPSON, JK
影响因子：
30.8
作者：
Mu, Jianbing;Awadalla, Philip;Su, Xin-zhuan
通讯作者：
Su, Xin-zhuan
影响因子：
30.8
作者：
Iafrate, AJ;Feuk, L;Lee, C
通讯作者：
Lee, C