Proceedings of the XLV Italian Society of Agricultural Genetics - SIGA Annual Congress

Salsomaggiore Terme, Italy - 26/29 September, 2001

ISBN 88-900622-1-5

Poster Abstract

A HALF MILLION CLONES BACTERIAL ARTIFICIAL CHROMOSOME (BAC) LIBRARY OF DURUM WHEAT

CENCI A.*, CHANTRET N.**, KONG X.-Y.***, GU Y.***, ANDERSON O.***, DUBCOVSKY J.****

* Dip. Biologia e Chimica Agroforestale e Ambientale, Università degli Studi di Bari, Via G. Amendola 165/a., 70126 Bari, Italia

alberto.cenci@agr.uniba.it

** Institut National de la Recherche Agronomique, Station d'Amelioration des Plantes, Domaine de la Motte, BP 29, 35653 Le Rheu Cedex, France

*** USDA Western Reg. Research Center, 800 Buchanan St., Albany, CA 94710, USA

**** Dept. Agronomy and Range Science, University of California, One Shields Avenue, Davis CA 95616-8515, USA

BAC library, durum wheat

A Triticum turgidum var.durum bacterial artificial chromosome (BAC) library was constructed. The genotype used for this library is a near isogenic line of Langdon carrying a 30 cM segment from the chromosome 6B of T. t. var. dicoccoides. More than half million of clones were organized in 1,344 384-well plates and 28 high-desity filters (with 18,432 double spotted clones each) were produced by using a Q-bot. With the aim to make possible PCR screening, plasmid DNA extraction was performed from bulked culture of bacterial strains of each 384-well plate. Based on the caracterization of 500 clones, the average size of the clones was 130Kb. Clones containing plastidial DNA and no insert (empty clones, false positives) percentages were estimated and they account for 0.34% and less than 1%, respectively. The genomic coverage of this library is 5X and the probability to recover any genomic sequence is bigger than 99%. Screening by filter hybridization with single- and low copy probes and PCR on plasmidic DNA (organized in bulks) were performed to assess the actual representativity of this BAC library. The results indicated a good correspondence between the observed and the expected data. Data about this library are available at: http://agronomy.ucdavis.edu/Dubcovsky/BAC-library/BAC_Langdon.htm