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.
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.
Jiménez
L V
LV
Center for Tropical Disease Research and Training, Department of Biological Sciences, University of Notre Dame, Notre Dame, IN 46556-5645, USA. David.W.Severson.1@nd.edu
Kang
B-K
BK
deBruyn
B
B
Lovin
D D
DD
Severson
D W
DW
D000330:Aedes / Q000235:genetics* / Q000469:parasitology*; D000818:Animals; D002874:Chromosome Mapping; D022202:Chromosomes, Artificial, Bacterial / Q000235:genetics*; D017931:DNA Primers; D016521:Electrophoresis, Gel, Pulsed-Field; D018274:Electroporation; D015723:Gene Library; D010961:Plasmodium / Q000472:pathogenicity; D040641:Quantitative Trait Loci / Q000235:genetics*; D016324:Sequence Tagged Sites
D017931:DNA Primers