Construction of a BAC library and generation of BAC end sequence-tagged connectors for genome sequencing of the African malaria mosquito Anopheles gambiae

被引:22
作者
Hong, YS
Hogan, JR
Wang, X
Sarkar, A
Sim, C
Loftus, BJ
Ren, C
Huff, ER
Carlile, JL
Black, K
Zhang, HB
Gardner, MJ
Collins, FH [1 ]
机构
[1] Univ Notre Dame, Dept Sci Biol, Ctr Trop Dis Res & Training, Notre Dame, IN 46556 USA
[2] Texas A&M Univ, Dept Soil & Crop Sci, College Stn, TX 77843 USA
[3] Texas A&M Univ, Inst Plant Genom & Biotechnol, College Stn, TX 77843 USA
[4] Inst Genome Res, Rockville, MD 20850 USA
关键词
Anopheles gambiae; Bacterial artificial chromosome (BAC) library; DNA sequencing; in situ hybridization;
D O I
10.1007/s00438-003-0813-7
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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), similar to2,000 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.
引用
收藏
页码:720 / 728
页数:9
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