Construction, characterization, and preliminary BAC-end sequencing analysis of a bacterial artificial chromosome library of white clover (Trifolium repens L.)

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dc.contributor.author Febrer, Melanie
dc.contributor.author Cheung, Foo
dc.contributor.author Town, Christopher D.
dc.contributor.author Cannon, Steven B.
dc.contributor.author Young, Nevin D.
dc.contributor.author Abberton, Michael T.
dc.contributor.author Jenkins, Glyn
dc.contributor.author Milbourne, Dan
dc.date.accessioned 2009-07-09T10:56:09Z
dc.date.available 2009-07-09T10:56:09Z
dc.date.issued 2007-04-01
dc.identifier.citation Febrer , M , Cheung , F , Town , C D , Cannon , S B , Young , N D , Abberton , M T , Jenkins , G & Milbourne , D 2007 , ' Construction, characterization, and preliminary BAC-end sequencing analysis of a bacterial artificial chromosome library of white clover (Trifolium repens L.) ' Genome , vol 50 , no. 4 , pp. 412-421 . en
dc.identifier.issn 1480-3321
dc.identifier.other PURE: 108918
dc.identifier.other dspace: 2160/2602
dc.identifier.uri http://hdl.handle.net/2160/2602
dc.description Febrer, M., Cheung, F., Town, C. D., Cannon, S. B., Young, N. D., Abberton, M. T., Jenkins, G., Milbourne, D. (2007). Construction, characterization, and preliminary BAC-end sequencing analysis of a bacterial artificial chromosome library of white clover (Trifolium repens L.). Genome, 50, (4), 412-421. Sponsorship: Biotechnology and Biological Sciences Research Council (BBSRC) en
dc.description.abstract White clover (Trifolium repens L.) is a forage legume widely used in combination with grass in pastures because of its ability to fix nitrogen. We have constructed a bacterial artificial chromosome (BAC) library of an advanced breeding line of white clover. The library contains 37 248 clones with an average insert size of approximately 85 kb, representing an approximate 3-fold coverage of the white clover genome based on an estimated genome size of 960 Mb. The BAC library was pooled and screened by polymerase chain reaction (PCR) amplification using both white clover microsatellites and PCR-based markers derived from Medicago truncatula, resulting in an average of 6 hits per marker; this supports the estimated 3-fold genome coverage in this allotetraploid species. PCR-based screening of 766 clones with a multiplex set of chloroplast primers showed that only 0.5% of BAC clones contained chloroplast-derived inserts. The library was further evaluated by sequencing both ends of 724 of the clover BACs. These were analysed with respect to their sequence content and their homology to the contents of a range of plant gene, expressed sequence tag, and repeat element databases. Forty-three microsatellites were discovered in the BAC-end sequences (BESs) and investigated as potential genetic markers in white clover. The BESs were also compared with the partially sequenced genome of the model legume M. truncatula with the specific intention of identifying putative comparative-tile BACs, which represent potential regions of microsynteny between the 2 species; 14 such BACs were discovered. The results suggest that a large-scale BAC-end sequencing strategy has the potential to anchor a significant proportion of the genome of white clover onto the gene-space sequence of M. truncatula. en
dc.format.extent 10 en
dc.language.iso eng
dc.relation.ispartof Genome en
dc.subject white clover en
dc.subject Trifolium repens en
dc.subject Medicago truncatula en
dc.subject legumes en
dc.subject comparative genomics en
dc.subject microsynteny en
dc.subject BAC library en
dc.subject BAC-end sequencing en
dc.subject microsatellite en
dc.title Construction, characterization, and preliminary BAC-end sequencing analysis of a bacterial artificial chromosome library of white clover (Trifolium repens L.) en
dc.type Text en
dc.type.publicationtype Article (Journal) en
dc.identifier.doi http://dx.doi.org/10.1139/G07-013
dc.contributor.institution Institute of Biological, Environmental and Rural Sciences en
dc.description.status Peer reviewed en


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