Protocol

Zebrafish

Volume 546 of the series Methods in Molecular Biology pp 69-84

Date:

Transient and Stable Transgenesis Using Tol2 Transposon Vectors

  • Hiroshi KikutaAffiliated withDivision of Molecular and Developmental Biology, National Institute of Genetics
  • , Koichi KawakamiAffiliated withDivision of Molecular and Developmental Biology, National Institute of GeneticsDepartment of Genetics, The Graduate University for Advanced Studies (SOKENDAI) Email author 

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Summary

Transgenesis is an important methodology for studying the function of genes and genomes in model plants and animals. For the model vertebrate zebrafish, methods using the Tol2 transposable element have been developed for this purpose. With these methods, the function of the transgene can be analyzed in both transient and stable transgenic fish. Recently, cis-sequences necessary for transposition of the Tol2 element were revealed. This enabled development of transposon vectors containing minimal DNA sequences that are easily manipulated. More recently, several transposon vectors containing the Gateway sequence were created and reported. These are useful because any foreign sequences can be cloned into a transposon vector fairly easily and rapidly. This chapter describes the features of these transposon vectors, and protocols to perform transgenesis in zebrafish.

Key words

Transposon Transposase Transposition Tol2 Transient transgenesis Stable transgenesis Microinjection Green fluorescent protein