Novel procedure of Trimethylpsoralen mutagenesis

[Hideki Ando]

Information on TMP mutagenesis procedure

Dear all,

We will report a novel procedure for efficient mutagenesis of zabrafish.
This method uses a DNA cross-linking agent 4,5',8-trimethylpsoralen
(TMP), which is known to frequently induce small deletions in
Escherichia coli and Caenorhabditis elegans. Our pilot screening
indicated that the TMP mutagenesis procedure was efficient. At present,
we are using 3 ng/ml TMP in final concentration to obtain mutants. The
high efficiency of the TMP mutagenesis will allow the isolation of a
significant number of zebrafish mutants in a single laboratory. TMP
induced small deletions by nucleotide excision and recombinational
repair of the interstrand DNA cross-links would be a useful marker for
the cloning of the mutated gene. In fact, we have successfully cloned a
zebrafish DNA fragment which is absent in a mutant. The procedure will
be detailed in the next issue of the Zebrafish Science Monitor. We hope
this method will contribute to zebrafish research community.

(Reference) Efficient mutagenesis of zebrafish by a DNA cross-linking
agent Neurosci. Lett. 244, 81-84. (1998)

Regards,
Hideki Ando

Hideki Ando, Ph.D.
Dapartment of Molecular Neurobiology and Pharmacology,
School of Medicine, University of Tokyo,
Hongo 7-3-1, Bunkyo-ku, Tokyo 113-0033, Japan
(Phone) 81-3-5689-7223
(FAX) 81-3-5689-7224
(E-mail) ykr2ando at m.u-tokyo.ac.jp