Analysis of extrachromosomal homologous recombination in cultured silkworm cells

Hiroaki Mon, Takahiro Kusakabe, Hisanori Bando, Katsura Kojima, Yutaka Kawaguchi, Katsumi Koga

Research output: Contribution to journalArticlepeer-review

9 Citations (Scopus)

Abstract

Double-strand breaks (DSBs) are potentially lethal lesions causing the loss of chromosomal information. Eukaryotic cells have evolved the error-free repair systems of DSBs by homologous recombination (HR) through gene conversion with or without crossing over. In this study, we have developed a rapid assay system for extrachromosomal HR events in the cultured silkworm BmN4 cells. When HR occurs within the disrupted luciferase gene, an enzymatically active luciferase is restored and expressed. Our results strongly suggest that error-prone single strand annealing (SSA) accounts for the majority of extrachromosomal recombination processes in the cells. However, upon the substrates which cannot be repaired through SSA, DSBs were efficiently repaired though gene conversion. The rapid and sensitive HR assay system developed in the present study is expected to be a powerful tool for the identification and analysis of HR-related genes in the silkworm.

Original languageEnglish
Pages (from-to)684-690
Number of pages7
JournalBiochemical and Biophysical Research Communications
Volume312
Issue number3
DOIs
Publication statusPublished - Dec 19 2003

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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