The CIC YAC library : Sizing of the clones and determination of clones carrying repeated DNA sequences

Francine Creusot, Eric Fouilloux and Michel Dron
Laboratoire de Phytopathologie Moléculaire, Institut de Biotechnologie des Plantes URA CNRS 1128, Université Paris-sud, 91405 Orsay Cedex France

Jacqueline Lafleuriel and Georges Picard
URA CNRS 1940, Biomove, Université Blaise-Pascal, 24 Avenue des Landais, 63177 Aubière Cedex France

Alain Billault, Denis Le Paslier and Daniel Cohen
Centre d'Etude du Polymorphisme Humain (CEPH), 27 rue Juliette Dodu, 75010 Paris France

Marie-Edith Chabouté, Andrée Durr, Jacqueline Fleck and Claude Gigot
Institut de Biologie Moléculaire des Plantes, Université Louis Pasteur, 12 rue du Général Zimmer, 67084 Strasbourg Cedex France

Christine Camilleri, Catherine Bellini, Michel Caboche and David Bouchez*
Laboratoire de Biologie Cellulaire, INRA, 78026 Versailles Cedex France

* author for correspondence (e-mail : bouchez@versailles.inra.fr).

Introduction

We have constructed a new Arabidopsis (ecotype Columbia) genomic library in Yeast Artificial Chromosomes : the CIC library (for CEPH/INRA/CNRS). The library consists of 1152 clones with an overall average insert size of 420 kb. The construction and characterization of the library are described elsewhere (Creusot et al., 1995). The CIC library is available to the community through the Arabidopsis Biological Resource Center (Ohio State University, USA).

Here we present the detailed data on size and content in repeated DNA for all the clones in the library. Detailed protocols and discussion of the results are presented in Creusot et al. (1995).

Material and methods

Briefly, yeast DNA plugs were prepared for all the 1152 clones in microtiter plates. The chromosomes were separated by PFGE CHEF electrophoresis and blotted onto nylon membranes. YACs were revealed by hybridization to a pBR322 probe. Their size were determined by comparison to yeast chromosomes (strain AB1380). The same membranes were used for hybridization with various probes corresponding to chloroplastic and mitochondrial DNA, or nuclear repeated DNA sequences (180 and 160 bp paracentromeric elements, 18S-25S rDNA, 5S rDNA, telomeric repeats).

Size and hybridization data of the CIC YAC library

No clone was found to hybridize to telomeric repeat or mitochondrial DNA probes. Only one hybridizes to the 160 bp element probe (CIC9E6).

The results concerning other probes are presented in a series of tables below. Each table shows the size and hybridization data of the CIC YAC library and is formatted in the same way. The table of information has been split for ease of access.

Column 1 (YAC #) : coordinates of the clones are according to the plate (1 to 12), rows (A to H) and columns (1 to 12).

Columns 2 to 6 : size of the YAC(s) (up to 5 YACs), estimated by comparison to yeast chromosomes (strain AB1380). Note that size data are missing for some of the clones. This may be due to problems during yeast culture and/or DNA plug preparation ; therefore some of these clones may actually contain YAC(s).

Hybridization : "+" indicates a positive hybridization signal. When two YACs are present, ">+", "<+", and "2+" indicate respectively hybridization with the larger, the smaller, or both YACs.

TOTAL : hybridization to total genomic DNA probe (including cpDNA) from Arabidopsis.

cpDNA : mixture of chloroplastic DNA probes from wheat, totalizing 60 kb of the organellar genome.

5S : Arabidopsis 5S rDNA probe (Campbell et al., 1992).

rDNA : 18S-25S rDNA intergenic spacer probe from Arabidopsis.

180 and 160 : respectively 180 bp and 160 bp paracentromeric element probes (Martinez-Zapater et al., 1986 ; Simoens et al., 1988).

Select the clones of interest and use the Netscape browser to view the tables. If your browser does not support viewing tables, please select from the second list below.

Tables

Text versions of Tables

References

Creusot, F., Fouilloux, E., Dron, M., Lafleuriel, J., Picard, G., Billault, A., Le Paslier, D., Cohen, D., Chabouté, M-E., Durr, A., Fleck, J., Gigot, C., Camilleri, C., Bellini, C., Caboche, M. and Bouchez, D. (1995) The CIC library : a large insert YAC library for genome mapping in Arabidopsis thaliana. Plant J. (in press)

Campbell, B.R., Song, Y., Posch, T.E., Cullis, C.A. and Town, C.D. (1992) Sequence and organization of 5S ribosomal RNA-encoding genes of Arabidopsis thaliana. Gene. 112, 225-228.

Martinez-Zapater, J.M., Estelle M.A. and Somerville, C.R. (1986) A highly repeated DNA sequence in Arabidopsis thaliana. Mol. Gen. Gen. 204, 417-423.

Simoens, C.R., Gielen, J., Van Montagu and Inzé, D. (1988) Characterization of highly repetitive sequences of Arabidopsis thaliana. Nucleic Acids Res. 16, 6753-6766.