Plasmodium falciparum:: A novel method for analyzing haplotypes in mixed infections
Plasmodium falciparum:: A novel method for analyzing haplotypes in mixed infections
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DOI：
10.1016/j.exppara.2006.09.003
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发表时间：
2007-03-01
影响因子：
2.1
通讯作者：
Sibley, Carol H.
中科院分区：
文献类型：
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作者：
Certain, Laura K.;Sibley, Carol H.
研究恶性疟原虫的群体遗传学对于了解耐药性的传播是必要的。然而，由于无法从包含多个寄生虫种群的患者样本中确定单倍型，这些研究受到了阻碍。因此，我们开发了一种分离混合感染中单个菌株进行遗传分析的方法。我们扩增了4号染色体6kb的片段，包括二氢叶酸还原酶基因和上游微卫星标记。该聚合酶链式反应产物通过重组插入到含有可选择标记和反选择标记的裂隙酵母穿梭质粒中。因为每个质粒只包含一个插入物，每个酵母菌落只包含一个质粒，所以各个菌株现在是分开的。我们分析了3D7、K1和DD2 DNA的混合物，并正确地识别了每种情况下的单倍型。(C)2006 Elsevier Inc.保留所有权利。
Studying the population genetics of Plasmodium falciparum is necessary for understanding the spread of drug resistance. However, these studies are hampered by the inability to determine haplotypes from patient samples that contain multiple parasite populations. Therefore, we have developed a method for separating for genetic analysis the individual strains in a mixed infection. We amplified a 6 kb region of chromosome 4, including the dihydrofolate reductase gene and upstream microsatellite markers. This PCR product was inserted by recombination into a gapped yeast shuttle plasmid containing both selectable and counter-select able markers. Because each plasmid contains only one insert and each yeast colony contains only one plasmid, the individual strains are now separate. We analyzed mixtures of 3D7, K1, and Dd2 DNA and correctly identified a haplotype in each case. (c) 2006 Elsevier Inc. All rights reserved.