TY - JOUR
T1 - Characterization of mutations induced by ethyl methanesulfonate, UV, and trimethylpsoralen in the nematode Caenorhabditis elegans
AU - Gengyo-Ando, Keiko
AU - Mitani, Shohei
N1 - Funding Information:
We thank Sachiko Noguchi for excellent technical assistance. We are grateful to Drs. Yuji Kohara, Yasumi Ohshima, Harumasa Okamoto, Asako Sugimoto, Masa-aki Muramatsu, and Hatsusi Shimizu for discussion. K.G.-A. is grateful to Dr. Masahiro Mu-kaida for his encouragement. Some strains used in this work were provided by Caenorhabditis Genetic Center, which is funded by the NIH National Center for Research Resources (NCRR). This work was supported partly by grants from Japanese Ministry of Education, Science, Sports and Culture, Japan Society for the Promotion of Science, and Japan Science and Technology Corporation to S.M.
PY - 2000/3/5
Y1 - 2000/3/5
N2 - The genome project of the nematode Caenorhabditis elegans is completed. It is important and useful to disrupt nematode genes to know their function. We treated wild-type animals with potential candidates for mutagens for reverse genetics, EMS (ethyl methanesulfonate), short-wavelength UV, and long-wavelength UV in the presence of TMP (trimethylpsoralen). We estimated forward mutation rates by counting the occurrence of a marker unc-22 mutation. We found that the forward mutation rate by TMP/UV could be comparable with EMS by improving the frequency one order higher than before. We next isolated mutants of another marker gene ben-1 and examined the probability for the deletion mutations by PCR and sequencing. Deletion mutations were found only by TMP/UV method, which suggested TMP/UV is the choice for deletion mutagenesis among these methods. As a pilot experiment, we could isolate actual deletion mutations at a much higher frequency than previously. (C) 2000 Academic Press.
AB - The genome project of the nematode Caenorhabditis elegans is completed. It is important and useful to disrupt nematode genes to know their function. We treated wild-type animals with potential candidates for mutagens for reverse genetics, EMS (ethyl methanesulfonate), short-wavelength UV, and long-wavelength UV in the presence of TMP (trimethylpsoralen). We estimated forward mutation rates by counting the occurrence of a marker unc-22 mutation. We found that the forward mutation rate by TMP/UV could be comparable with EMS by improving the frequency one order higher than before. We next isolated mutants of another marker gene ben-1 and examined the probability for the deletion mutations by PCR and sequencing. Deletion mutations were found only by TMP/UV method, which suggested TMP/UV is the choice for deletion mutagenesis among these methods. As a pilot experiment, we could isolate actual deletion mutations at a much higher frequency than previously. (C) 2000 Academic Press.
KW - Caenorhabditis elegans
KW - Deletion mutant
KW - Reverse genetics
KW - Trimethylpsoralen
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U2 - 10.1006/bbrc.2000.2260
DO - 10.1006/bbrc.2000.2260
M3 - Article
C2 - 10694478
AN - SCOPUS:0034607118
VL - 269
SP - 64
EP - 69
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
SN - 0006-291X
IS - 1
ER -