Construction of a rice bacterial artificial chromosome library and identification of clones linked to the Xa‐21 disease resistance locus

Authors:
  • PRPamela C. Ronald
    Pamela C. Ronald
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Abstract
Summary: A bacterial artificial chromosome (BAC) library consisting of 11 000 clones with an average DNA insert size of 125 kb was constructed from rice nuclear DNA. The BAC clones were stable in E. coli after 100 generations of serial growth. Transformation of the BAC clones by electroporation into E. coli was highly efficient and increased with decreasing size of the DNA inserts. The library was evaluated for the presence of organellar, repeated, and telomeric sequences. A very low percentage (Arabidopsis telomere repeat. Sixteen BACs hybridized with the AA genome‐specific repetitive sequence pOs48. Twelve clones were isolated that hybridized with three DNA markers linked to the Xa‐21 disease resistance locus. The results indicate that the BAC system can be used to clone and manipulate large pieces of plant DNA efficiently.