Characterization of an Aedes aegypti bacterial artificial chromosome (BAC) library and chromosomal assignment of BAC clones for physical mapping quantitative trait loci that influence Plasmodium susceptibility
Characterization of an Aedes aegypti bacterial artificial chromosome (BAC) library and chromosomal assignment of BAC clones for physical mapping quantitative trait loci that influence Plasmodium susceptibility
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DOI：
10.1046/j.0962-1075.2004.00456.x
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发表时间：
2004-02-01
影响因子：
2.6
通讯作者：
Severson, DW
中科院分区：
文献类型：
--
作者：
Jiménez, LV;Kang, B;Severson, DW
以前的研究已经证实了蚊子对疟原虫易感性的遗传基础。在这里，我们描述了我们为黄热病蚊子埃及伊蚊的细菌人工染色体基因组文库所做的工作，并鉴定了包含定义鸡疟原虫易感性的数量性状基因座(QTL)的遗传标记的BAC克隆。该文库(NDL)是从Ae.埃及伊蚊利物浦株，由排列在384孔微孔板上的50 304个克隆组成。我们使用针对106个遗传标记(作为序列标记位点或STS)的寡核苷酸引物对NDL文库进行了筛选。每个STS鉴定出1至13个独立克隆，平均3.3个克隆。平均插入大小为122kb，因此NDL文库提供了大约7.87倍的基因组覆盖率。NDL文库的可获得性将极大地促进物理作图工作，包括对疟疾易感性等感兴趣特征的QTL的定位克隆，以及用于全基因组序列的确定和组装。
Previous studies have confirmed a genetic basis for susceptibility of mosquitoes to Plasmodium parasites. Here we describe our efforts to characterize a bacterial artificial chromosome genomic library for the yellow fever mosquito, Aedes aegypti, and to identify BAC clones containing genetic markers that define quantitative trait loci (QTL) for Plasmodium gallinaceum susceptibility. This library (NDL) was prepared from the Ae. aegypti Liverpool strain and consists of 50 304 clones arrayed in 384-well microplates. We used PCR analysis with oligonucleotide primer pairs specific to 106 genetic markers (as sequence-tagged sites or STS) to screen the NDL library. Each STS identified between one and thirteen independent clones with an average of 3.3 clones. The average insert size was 122 kb and therefore the NDL library provides approximately 7.87-fold genome coverage. The availability of the NDL library should greatly facilitate physical mapping efforts, including positional cloning of QTL for traits of interest such as Plasmodium susceptibility and for whole genome sequence determination and assembly.