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New genomic resources for the honey bee
(Apis mellifera L.): development of a
deep-coverage BAC library and a
preliminary STC database
J.P. Tomkins1, M. Luo2, G.C. Fang1, D. Main1, J.L. Goicoechea2,
M. Atkins1, D.A. Frisch3, R.E. Page4, E. Guzmán-Novoa5, Y. Yu2,
G. Hunt6 and R.A. Wing2
1Clemson University Genomics Institute, Room 100 Jordan Hall, Clemson, SC 29634, USA
2Arizona Genomics Institute, 303 Forbes Hall, Tucson, AZ 85721, USA
3Genome Center of Wisconsin, 445 Henry Mall, Genetics Building B19, Madison, WI 53706, USA
4Department of Entomology, University of California, Davis, CA 95616, USA
5INIFAP/SAGAR, 29B Santa Cruz, Faracc., Las Hdas., Metepéc, 52140, Edo. de Mex., Mexico
6Department of Entomology, Purdue University, 1158 Smith Hall, West Lafayette, IN 47907, USA
Corresponding author: J.P. Tomkins
E-mail: [email protected]
Genet. Mol. Res. 1 (4): 306-316 (2002)
Received September 23, 2002
Published November 22, 2002

ABSTRACT. We have constructed a bacterial artificial chromosome (BAC) library for a European honey bee strain using the cloning enzyme HindIII in order to develop resources for structural genomics research. The library contains 36,864 clones (ninety-six 384-well plates). A random sampling of 247 clones indicated an average insert size of 113 kb (range = 27 to 213 kb) and 2% empty vectors. Based on an estimated genome size of 270 Mb, this library provides approximately 15 haploid genome equivalents, allowing >99% probability of recovering any specific sequence of interest. High-density colony filters were gridded robotically using a Genetix Q-BOT in a 4 x 4 double-spotted array on 22.5-cm2 filters. Screening of the library with four mapped honey bee genomic clones and two bee cDNA probes identified an average of 21 positive signals per probe, with a range of 7-38 positive signals per probe. An additional screening was performed with nine aphid gene fragments and one Drosophila gene fragment resulting in seven of the nine aphid probes and the Drosophila probe producing positive signals with a range of 1 to 122 positive signals per probe (average of 45). To evaluate the utility of the library for sequence tagged connector analysis, 1152 BAC clones were end sequenced in both forward and reverse directions, giving a total of 2061 successful reads of high quality. End sequences were queried against SWISS-PROT, insect genomic sequence GSS, insect EST, and insect transposable element databases. Results in spreadsheet format from these searches are publicly available at the Clemson University Genomics Institute (CUGI) website in a searchable format (http://www.genome.clemson.edu/projects/stc/bee/AM__Ba/).

Key words: Bacterial artificial chromosome, BAC, Honey bee, Comparative genomics, BAC end sequencing, STC

 

 

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