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Open Access Short Report

Enhancing genome investigations in the mosquito Culex quinquefasciatus via BAC library construction and characterization

Paul V Hickner1, Becky deBruyn1, Diane D Lovin1, Akio Mori1, Christopher A Saski2 and David W Severson1*

Author Affiliations

1 Eck Institute for Global Health, Department of Biological Sciences, University of Notre Dame, Notre Dame, IN 46556, USA

2 Clemson University Genomics Institute, Clemson, SC 29634, USA

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BMC Research Notes 2011, 4:358  doi:10.1186/1756-0500-4-358

Published: 13 September 2011

Abstract

Background

Culex quinquefasciatus (Say) is a major species in the Culex pipiens complex and an important vector for several human pathogens including West Nile virus and parasitic filarial nematodes causing lymphatic filariasis. It is common throughout tropical and subtropical regions and is among the most geographically widespread mosquito species. Although the complete genome sequence is now available, additional genomic tools are needed to improve the sequence assembly.

Findings

We constructed a bacterial artificial chromosome (BAC) library using the pIndigoBAC536 vector and HindIII partially digested DNA isolated from Cx. quinquefasciatus pupae, Johannesburg strain (NDJ). Insert size was estimated by NotI digestion and pulsed-field gel electrophoresis of 82 randomly selected clones. To estimate genome coverage, each 384-well plate was pooled for screening with 29 simple sequence repeat (SSR) and five gene markers. The NDJ library consists of 55,296 clones arrayed in 144 384-well microplates. Fragment insert size ranged from 50 to 190 kb in length (mean = 106 kb). Based on a mean insert size of 106 kb and a genome size of 579 Mbp, the BAC library provides ~10.1-fold coverage of the Cx. quinquefasciatus genome. PCR screening of BAC DNA plate pools for SSR loci from the genetic linkage map and for four genes associated with reproductive diapause in Culex pipiens resulted in a mean of 9.0 positive plate pools per locus.

Conclusion

The NDJ library represents an excellent resource for genome assembly enhancement and characterization in Culex pipiens complex mosquitoes.

Keywords:
Bacterial Artificial Chromosome; Culicidae; Culex pipiens/quinquefasciatus; Lymphatic filariasis; West Nile virus; Wuchereria bancrofti