Construction and characterization of a bacterial artificial chromosome library of peach

Q. Wang, K. Zhang, X. Qu, J. Jia, J. Shi, D. Jin, B. Wang

Research output: Contribution to journalArticlepeer-review

25 Scopus citations

Abstract

A peach [Prunus persica (L.) Batch] bacterial artificial chromosome (BAC) library of var. Jingyu was constructed. Jingyu is a traditional variety, that displays many of the important agronomic characters of stone fruits. Since peach leaves are rich in polysaccharides, high-molecular-weight (HMW) DNA was extracted from leaf nuclei using a protocol adapted to peach. The HMW DNA embedded in agarose plugs was partially digested by HindIII. After size-selection by pulsed field gel electrophoresis, the selected DNA fragments were ligated to pBeloBACll and transformed into E. coli DH10B cells by electroporation. In total 20,736 recombinant clones were obtained. The BAC library has an average insert size of 95 kb and represents approximately 6.7 peach haploid genome equivalents. The BAC clones were stable in E. coli cell after 100 generations. The lack of hybridization to chloroplast and mitochondrial genes demonstrated that the library is predominantly composed of nuclear DNA. The library was screened with two molecular markers, W4 and P20, that are linked to white flesh and nectarine genes of peach, respectively. Ten positive clones were detected. Their fingerprints will be used to determine clone relationships and assemble contigs. This library should be well-suited for the map-based cloning of peach genes and genome physical mapping.

Original languageEnglish
Pages (from-to)1174-1179
Number of pages6
JournalTheoretical And Applied Genetics
Volume103
Issue number8
DOIs
StatePublished - 2001

Keywords

  • Bacterial artificial chromosome
  • High-molecular-weight DNA
  • Prunus persica (peach)

ASJC Scopus subject areas

  • Biotechnology
  • Agronomy and Crop Science
  • Genetics

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