Instability of bacterial artificial chromosome (BAC) clones containing tandemly repeated DNA sequences

J. Song, F. Dong, J. W. Lilly, R. M. Stupar, J. Jiang

Research output: Contribution to journalArticlepeer-review

34 Scopus citations

Abstract

The cloning and propagation of large DNA fragments as bacterial artificial chromosomes (BACs) has become a valuable technique in genome research. BAC clones are highly stable in the host, Escherichia coli, a major advantage over yeast artificial chromosomes (YACs) in which recombination-induced instability is a major drawback. Here we report that BAC clones containing tandemly repeated DNA elements are not stable and can undergo drastic deletions during routine library maintenance and DNA preparation. Instability was observed in three BAC clones from sorghum, rice, and potato, each containing distinct tandem repeats. As many as 46% and 74% of the single colonies derived from a rice BAC clone containing 5S ribosomal RNA genes had insert deletions after 24 and 120 h of growth, respectively. We also demonstrated that BAC insert rearrangement can occur in the early stage of library construction and duplication. Thus, a minimum growth approach may not avoid the instability problem of such clones. The impact of BAC instability on genome research is discussed.

Original languageEnglish (US)
Pages (from-to)463-469
Number of pages7
JournalGenome
Volume44
Issue number3
DOIs
StatePublished - Jan 1 2001

Keywords

  • Genome research
  • Large insert DNA library
  • Repetitive DNA

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