Construction and characterization of a bacterial artificial chromosome (BAC) library for the A genome of wheat

D. Lijavetzky, G. Muzzi, T. Wicker, B. Keller, Rod A Wing, J. Dubcovsky

Research output: Contribution to journalArticle

118 Citations (Scopus)

Abstract

A genomic bacterial artificial chromosome (BAC) library of the A genome of wheat has been constructed. Triticum monococcum accession DV92 was selected for this purpose because it is a cultivated diploid wheat and one of the parental lines used in the construction of a saturated genetic map. Leaves from this accession were used to isolate high-molecular-weight DNA from nuclei. This DNA was partially digested with restriction enzyme Hind III, subjected to double size selection, electroeluted and cloned into the pINDIGO451 BAC vector. The library consists of 276 480 clones with an average insert size of 115 kb. Excluding the 1.33% of empty clones and 0.14% of clones with chloroplast DNA, the coverage of this library is 5.6 genome equivalents. With this genome coverage the probability of having any DNA sequence represented in this library is higher than 99.6%. Clones were sorted in 720 384-well plates and blotted onto 15 high-density filters. High-density filters were screened with several single or low-copy clones and five positive BAC clones were selected for further analysis. Since most of the T. monococcum BAC ends included repetitive sequences, a modification was introduced into the classical end-isolation procedure to select low copy sequences for chromosome walking.

Original languageEnglish (US)
Pages (from-to)1176-1182
Number of pages7
JournalGenome
Volume42
Issue number6
StatePublished - 1999
Externally publishedYes

Fingerprint

Bacterial Artificial Chromosomes
bacterial artificial chromosomes
Genomic Library
Triticum
Clone Cells
clones
wheat
genome
Triticum monococcum
Libraries
Chromosome Walking
chromosome walking
Genome
Chloroplast DNA
repetitive sequences
DNA libraries
Nucleic Acid Repetitive Sequences
DNA
Diploidy
chloroplast DNA

Keywords

  • BAC library
  • Bacterial artificial chromosome
  • Triticum monococcum
  • Wheat

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biotechnology
  • Genetics
  • Genetics(clinical)

Cite this

Lijavetzky, D., Muzzi, G., Wicker, T., Keller, B., Wing, R. A., & Dubcovsky, J. (1999). Construction and characterization of a bacterial artificial chromosome (BAC) library for the A genome of wheat. Genome, 42(6), 1176-1182.

Construction and characterization of a bacterial artificial chromosome (BAC) library for the A genome of wheat. / Lijavetzky, D.; Muzzi, G.; Wicker, T.; Keller, B.; Wing, Rod A; Dubcovsky, J.

In: Genome, Vol. 42, No. 6, 1999, p. 1176-1182.

Research output: Contribution to journalArticle

Lijavetzky, D, Muzzi, G, Wicker, T, Keller, B, Wing, RA & Dubcovsky, J 1999, 'Construction and characterization of a bacterial artificial chromosome (BAC) library for the A genome of wheat', Genome, vol. 42, no. 6, pp. 1176-1182.
Lijavetzky, D. ; Muzzi, G. ; Wicker, T. ; Keller, B. ; Wing, Rod A ; Dubcovsky, J. / Construction and characterization of a bacterial artificial chromosome (BAC) library for the A genome of wheat. In: Genome. 1999 ; Vol. 42, No. 6. pp. 1176-1182.
@article{8bbb73666a744b87aa97cdfc61f5ce06,
title = "Construction and characterization of a bacterial artificial chromosome (BAC) library for the A genome of wheat",
abstract = "A genomic bacterial artificial chromosome (BAC) library of the A genome of wheat has been constructed. Triticum monococcum accession DV92 was selected for this purpose because it is a cultivated diploid wheat and one of the parental lines used in the construction of a saturated genetic map. Leaves from this accession were used to isolate high-molecular-weight DNA from nuclei. This DNA was partially digested with restriction enzyme Hind III, subjected to double size selection, electroeluted and cloned into the pINDIGO451 BAC vector. The library consists of 276 480 clones with an average insert size of 115 kb. Excluding the 1.33{\%} of empty clones and 0.14{\%} of clones with chloroplast DNA, the coverage of this library is 5.6 genome equivalents. With this genome coverage the probability of having any DNA sequence represented in this library is higher than 99.6{\%}. Clones were sorted in 720 384-well plates and blotted onto 15 high-density filters. High-density filters were screened with several single or low-copy clones and five positive BAC clones were selected for further analysis. Since most of the T. monococcum BAC ends included repetitive sequences, a modification was introduced into the classical end-isolation procedure to select low copy sequences for chromosome walking.",
keywords = "BAC library, Bacterial artificial chromosome, Triticum monococcum, Wheat",
author = "D. Lijavetzky and G. Muzzi and T. Wicker and B. Keller and Wing, {Rod A} and J. Dubcovsky",
year = "1999",
language = "English (US)",
volume = "42",
pages = "1176--1182",
journal = "Genome / National Research Council Canada = Genome / Conseil national de recherches Canada",
issn = "0831-2796",
publisher = "National Research Council of Canada",
number = "6",

}

TY - JOUR

T1 - Construction and characterization of a bacterial artificial chromosome (BAC) library for the A genome of wheat

AU - Lijavetzky, D.

AU - Muzzi, G.

AU - Wicker, T.

AU - Keller, B.

AU - Wing, Rod A

AU - Dubcovsky, J.

PY - 1999

Y1 - 1999

N2 - A genomic bacterial artificial chromosome (BAC) library of the A genome of wheat has been constructed. Triticum monococcum accession DV92 was selected for this purpose because it is a cultivated diploid wheat and one of the parental lines used in the construction of a saturated genetic map. Leaves from this accession were used to isolate high-molecular-weight DNA from nuclei. This DNA was partially digested with restriction enzyme Hind III, subjected to double size selection, electroeluted and cloned into the pINDIGO451 BAC vector. The library consists of 276 480 clones with an average insert size of 115 kb. Excluding the 1.33% of empty clones and 0.14% of clones with chloroplast DNA, the coverage of this library is 5.6 genome equivalents. With this genome coverage the probability of having any DNA sequence represented in this library is higher than 99.6%. Clones were sorted in 720 384-well plates and blotted onto 15 high-density filters. High-density filters were screened with several single or low-copy clones and five positive BAC clones were selected for further analysis. Since most of the T. monococcum BAC ends included repetitive sequences, a modification was introduced into the classical end-isolation procedure to select low copy sequences for chromosome walking.

AB - A genomic bacterial artificial chromosome (BAC) library of the A genome of wheat has been constructed. Triticum monococcum accession DV92 was selected for this purpose because it is a cultivated diploid wheat and one of the parental lines used in the construction of a saturated genetic map. Leaves from this accession were used to isolate high-molecular-weight DNA from nuclei. This DNA was partially digested with restriction enzyme Hind III, subjected to double size selection, electroeluted and cloned into the pINDIGO451 BAC vector. The library consists of 276 480 clones with an average insert size of 115 kb. Excluding the 1.33% of empty clones and 0.14% of clones with chloroplast DNA, the coverage of this library is 5.6 genome equivalents. With this genome coverage the probability of having any DNA sequence represented in this library is higher than 99.6%. Clones were sorted in 720 384-well plates and blotted onto 15 high-density filters. High-density filters were screened with several single or low-copy clones and five positive BAC clones were selected for further analysis. Since most of the T. monococcum BAC ends included repetitive sequences, a modification was introduced into the classical end-isolation procedure to select low copy sequences for chromosome walking.

KW - BAC library

KW - Bacterial artificial chromosome

KW - Triticum monococcum

KW - Wheat

UR - http://www.scopus.com/inward/record.url?scp=0033493289&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0033493289&partnerID=8YFLogxK

M3 - Article

VL - 42

SP - 1176

EP - 1182

JO - Genome / National Research Council Canada = Genome / Conseil national de recherches Canada

JF - Genome / National Research Council Canada = Genome / Conseil national de recherches Canada

SN - 0831-2796

IS - 6

ER -