Construction and characterization of a common bean bacterial artificial chromosome library

Wim Vanhouten, Sally MacKenzie

Research output: Contribution to journalArticle

31 Scopus citations

Abstract

We have constructed a common bean (Phaseolus vulgaris L.) bacterial artificial chromosome (BAC) library consisting of 33 792 clones and an estimated 3- to 5-fold coverage of the common bean genome. Leaf nuclei were used as the source for high-molecular-weight DNA, and an endonuclease/methylase competition assay was employed to partially cleave the DNA. The library was screened with a number of nuclear and mitochondrial probes. Each nuclear probe identified at least two BACs with an average insert size of ca. 100 kb. Only 26 clones were identified after hybridizing with mitochondrial probes, indicating contamination with organellar sequences is low. Numerous clones could be identified after screening the library with two repetitive probes flanking the nuclear fertility restorer Fr. Intriguingly, 12 clones appeared to hybridize to both markers, and restriction analysis of these clones revealed that they can be assembled into maximally four contigs, suggesting that these repetitive probes may be useful for the physical mapping of the Fr locus.

Original languageEnglish (US)
Pages (from-to)977-983
Number of pages7
JournalPlant Molecular Biology
Volume40
Issue number6
DOIs
StatePublished - Aug 1 1999

Keywords

  • BAC library
  • Common bean
  • Fertility restorer
  • Legume genomics

ASJC Scopus subject areas

  • Agronomy and Crop Science
  • Genetics
  • Plant Science

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