Hong, Y. S., Hogan, J. R., Wang, X., Sarkar, A., Sim, C., Loftus, B. J., Ren, C., Huff, E. R., Carlile, J. L., Black, K., Zhang, H. B., Gardner, M. J., Collins, F. H.
Construction of a BAC Library and Generation of BAC End Sequence-tagged Connectors for Genome Sequencing of the African Malaria Mosquito Anopheles gambiae
Mol Genet Genomics. 2003 Mar 01; 268(6): 720-8.
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 whitelocus 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. gambiaegenome and for linking the assembled sequence to the three polytene chromosomes. This ND-TAM library is now publicly available at both www.malaria.mr4.org/mr4pages/index.html/ and hbz.tamu.edu, providing a valuable resource to the mosquito research community.