Construction and characterization of a Magnaporthe grisea bacterial artificial chromosome library

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

doi:10.1006/fgbi.1996.0042

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 journal › Article › peer-review

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 language	English (US)
Pages (from-to)	280-288
Number of pages	9
Journal	Fungal Genetics and Biology
Volume	20
Issue number	4
DOIs	https://doi.org/10.1006/fgbi.1996.0042
State	Published - Dec 1996

Keywords

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

ASJC Scopus subject areas

Microbiology
Genetics

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10.1006/fgbi.1996.0042

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title = "Construction and characterization of a Magnaporthe grisea bacterial artificial chromosome library",

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.",

keywords = "Bacterial artificial chromosome (BAC), Magnaporthe grisea, Map-based cloning, Rice blast",

author = "Diaz-Perez, {Silvia V.} and Crouch, {V. Wayne} and Orbach, {Marc J.}",

note = "Funding Information: We thank H. Shizuya for providing the pBeloBAC11 vector. We are grateful to B. Birren for providing unpublished protocols and advice about BAC library construction. Thanks to M. A. Mandel for assistance and U. Gunawardena and M. A. Mandel for critical reading of the manuscript. This work was supported by the USDA NRICGP (Grant 92-37303-7794).",

year = "1996",

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doi = "10.1006/fgbi.1996.0042",

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AU - Orbach, Marc J.

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PY - 1996/12

Y1 - 1996/12

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KW - Rice blast

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Construction and characterization of a Magnaporthe grisea bacterial artificial chromosome library

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Keywords

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