Journal article
New genomic resources for the honey bee(Apis mellifera L.): development of a deep-coverage BAC library and a preliminary STC database
Genetics and molecular research, Vol.1(4), pp.306-316
12/31/2002
Handle:
https://hdl.handle.net/2376/108283
PMID: 14963821
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/).
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Details
- Title
- New genomic resources for the honey bee(Apis mellifera L.): development of a deep-coverage BAC library and a preliminary STC database
- Creators
- J P Tomkins - Clemson University Genomics Institute, Room 100 Jordan Hall, Clemson, SC 29634, USA. jtmkns@CLEMSON.EDUM LuoG C FangD MainJ L GoicoecheaM AtkinsD A FrischR E PageE Guzmán-NovoaY YuG HuntR A Wing
- Publication Details
- Genetics and molecular research, Vol.1(4), pp.306-316
- Academic Unit
- Horticulture, Department of
- Publisher
- Brazil
- Identifiers
- 99900547141501842
- Language
- English
- Resource Type
- Journal article