Construction of a BAC library and generation of BAC end sequence-tagged connectors for genome sequencing of the African malaria mosquito Anopheles gambiae.
A Bacterial Artificial Chromosome (BAC) genomic DNA library of Anopheles gambiae, the major human malaria vector in sub-Saharan Africa, was constructed and characterized. This library (ND-TAM) is composed of 30,720 BAC clones in eighty 384-well plates. The estimated average insert size of the library is 133 kb, with an overall genome coverage of approximately 14-fold. The ends of approximately two-thirds of the clones in the library were sequenced, yielding 32,340 pair-mate ends. A statistical analysis (G-test) of the results of PCR screening of the library indicated a random distribution of BACs in the genome, although one gap encompassing the white locus on the X-chromosome was identified. Furthermore, combined with another previously constructed BAC library (ND-1), ~2,000 BACs have been physically mapped by polytene chromosomal in situ hybridization. These BAC end pair mates and physically mapped BACs have been useful for both the assembly of a fully sequenced A. gambiae genome and for linking the assembled sequence to the three polytene chromosomes. This ND-TAM library is now publicly available at both http://www.malaria.mr4.org/mr4pages/index.html/ and http://hbz.tamu.edu/, providing a valuable resource to the mosquito research community.
Hong
Y S
YS
Center for Tropical Disease Research and Training, Department of Biological Sciences, University of Notre Dame, IN 46556, USA.
Hogan
J R
JR
Wang
X
X
Sarkar
A
A
Sim
C
C
Loftus
B J
BJ
Ren
C
C
Huff
E R
ER
Carlile
J L
JL
Black
K
K
Zhang
H-B
HB
Gardner
M J
MJ
Collins
F H
FH
D000818:Animals; D000852:Anopheles / Q000235:genetics* / Q000469:parasitology; D022202:Chromosomes, Artificial, Bacterial / Q000235:genetics*; D015723:Gene Library; D017344:Genes, Insect; D016678:Genome; D006801:Humans; D017403:In Situ Hybridization; D007303:Insect Vectors / Q000235:genetics / Q000469:parasitology; D008288:Malaria / Q000635:transmission; D016133:Polymerase Chain Reaction; D016324:Sequence Tagged Sites