tailieunhanh - Báo cáo lâm nghiệp: "Toward a Pinus pinaster bacterial artificial chromosome library"

Tuyển tập các báo cáo nghiên cứu về lâm nghiệp được đăng trên tạp chí lâm nghiệp Original article đề tài: Toward a Pinus pinaster bacterial artificial chromosome library. | Ann. For. Sci. 64 2007 855-864 INRA EDP Sciences 2007 DOI forest 2007060 Available online at Original article Toward a Pinus pinaster bacterial artificial chromosome library Rocío Bautista David P. VILLALOBOS Sara Diaz-Moreno Francisco R. Canton Francisco M. Canovas M. Gonzalo Claros Departamento de Biología Molecular y Bioquímica Facultad de Ciencias Instituto Andaluz de Biotecnología Universidad de Malaga 29071 Malaga Spain Received 11 October 2006 accepted 7 February 2007 Abstract - Conifers are of great economic and ecological importance but little is known concerning their genomic organization. This study is an attempt to obtain high-quality high-molecular-weight DNA from Pinus pinaster cotyledons and the construction of a pine BAC library. The preparation incorporates modifications like low centrifugation speeds increase of EDTA concentration for plug maintenance use of DNase inhibitors to reduce DNA degradation use of polyvinylpyrrolidone and ascorbate to avoid secondary metabolites and a brief electrophoresis of the plugs prior to their use. A total of 72 192 clones with an average insert size of 107 kb which represents an equivalent of 11X pine haploid genomes were obtained. The proportions of clones lacking inserts or containing chloroplast DNA are both approximately . The library was screened with cDNA probes for seven genes and two clones containing Fd-GOGAT sequences were found one of them seemingly functional. Ongoing projects aimed at constructing a pine bacterial artificial chromosome library may benefit from the methods described here. BAC library glutamate synthase high molecular weight DNA pine Résumé - Construction d une banque BAC pour le pin maritime Pinus pinaster . Les conifères présentent un intérêt économique et écologique de premier plan mais restent très mal connus du point de vue de l organisation de leur génome. Cette étude présente une tentative réussie de construction d une banque BAC de séquences d .