TY - JOUR
T1 - Analysis of extrachromosomal homologous recombination in cultured silkworm cells
AU - Mon, Hiroaki
AU - Kusakabe, Takahiro
AU - Bando, Hisanori
AU - Kojima, Katsura
AU - Kawaguchi, Yutaka
AU - Koga, Katsumi
N1 - Funding Information:
This work was supported in part by Grant Nos. 14704010 and 14656024, and the National Bioresource Project, from the Ministry of Education, Science and Culture of Japan.
PY - 2003/12/19
Y1 - 2003/12/19
N2 - 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.
AB - 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.
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U2 - 10.1016/j.bbrc.2003.10.169
DO - 10.1016/j.bbrc.2003.10.169
M3 - Article
C2 - 14680819
AN - SCOPUS:0344552852
SN - 0006-291X
VL - 312
SP - 684
EP - 690
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
IS - 3
ER -