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Toward a Pinus pinaster bacterial artificial chromosome library
University of Malaga.
University of Malaga.
University of Malaga.ORCID-id: 0000-0002-3369-2440
University of Malaga.
Vise andre og tillknytning
2007 (engelsk)Inngår i: Annals of Forest Science, ISSN 1286-4560, E-ISSN 1297-966X, Vol. 64, nr 8, s. 855-864Artikkel i tidsskrift (Fagfellevurdert) Published
Abstract [en]

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 1.6%. 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 pinebacterial artificial chromosome library may benefit from the methods described here.

sted, utgiver, år, opplag, sider
2007. Vol. 64, nr 8, s. 855-864
Emneord [en]
BAC, Glutamate synthase, High molecular weight DNA, Library, Pine
HSV kategori
Identifikatorer
URN: urn:nbn:se:kth:diva-84671DOI: 10.1051/forest:2007060ISI: 000251157600008OAI: oai:DiVA.org:kth-84671DiVA, id: diva2:499561
Merknad
QC 20120223Tilgjengelig fra: 2012-02-13 Laget: 2012-02-13 Sist oppdatert: 2017-12-07bibliografisk kontrollert

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Forlagets fullteksthttp://dx.doi.org/10.1051/forest:2007060

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Diaz-Moreno, Sara M

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