Construction and characterization of a Magnaporthe grisea bacterial artificial chromosome library

Silvia V. Diaz-Perez, V. Wayne Crouch, Marc J. Orbach

Research output: Contribution to journalArticle

17 Scopus citations

Abstract

A bacterial artificial chromosome (BAC) library of Magnaporthe grisea containing 4128 clones with an average insert size of 66-kb has been constructed. This library represents seven genome equivalents of M. grisea and has been demonstrated to be representative of the genome by screening for the presence of several single-copy genes and DNA markers. The utility of the library for use in map-based cloning projects was shown by the spanning of a nine-cosmid, 207-kb DNA contig with only 3 BAC clones. In addition, using a lys1-3 auxotroph, we have shown that BAC clones at least 113 kb can be transformed into M. grisea to screen for complementation of mutations. Thus, BACs isolated in chromosome walks can be rapidly screened for the presence of the sought after gene. The ease of construction of BAC libraries and of isolation and manipulation of BAC clones makes the BAC system an ideal one for physical analyses of fungal genomes.

Original languageEnglish (US)
Pages (from-to)280-288
Number of pages9
JournalFungal Genetics and Biology
Volume20
Issue number4
DOIs
StatePublished - Dec 1996

Keywords

  • Bacterial artificial chromosome (BAC)
  • Magnaporthe grisea
  • Map-based cloning
  • Rice blast

ASJC Scopus subject areas

  • Microbiology
  • Genetics

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