Generation of transgenic medaka using modified bacterial artificial chromosome
Version of Record online: 16 APR 2008
© 2008 The Authors. Journal compilation © 2008 Japanese Society of Developmental Biologists
Development, Growth & Differentiation
Special Issue: Innovative techniques for the study of development
Volume 50, Issue 6, pages 415–419, August 2008
How to Cite
Nakamura, S., Saito, D. and Tanaka, M. (2008), Generation of transgenic medaka using modified bacterial artificial chromosome. Development, Growth & Differentiation, 50: 415–419. doi: 10.1111/j.1440-169X.2008.01027.x
- Issue online: 4 JUL 2008
- Version of Record online: 16 APR 2008
- Received 5 February 2008; revised 8 February 2008; accepted 27 February 2008.
The availability of bacterial artificial chromosome (BAC) offers a good genomic platform for a targeted integration of an exogenous gene by a homologous recombination system in Escherichia coli. In combination with microinjection technology, this system allows for the analysis of various aspects of biological phenomena occurring in vivo using Japanese medaka fish (Oryzias latipes). Here we describe a streamlined procedure for selecting BAC clones based on the medaka University of Tokyo genome browser (UTGB), followed by rapid modification with enhanced green fluorescent protein (EGFP) or DsRed fragments for transgenic analysis in medaka. Experimental procedures for BAC DNA preparation, microinjection of medaka embryos and screening of resulting transgenic medaka carrying EGFP/DsRed modified BAC clones are also described.