使用tol2转座子载体的瞬时和稳定转基因技术。

Transient and stable transgenesis using tol2 transposon vectors.

作者信息

Kikuta Hiroshi, Kawakami Koichi

机构信息

Division of Molecular and Developmental Biology, National Institute of Genetics, 1111 Yata, Mishima, Shizuoka, 411-8540, Japan.

出版信息

Methods Mol Biol. 2009;546:69-84. doi: 10.1007/978-1-60327-977-2_5.

DOI:10.1007/978-1-60327-977-2_5

PMID:19378098

Abstract

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.

摘要

转基因技术是研究模式动植物中基因和基因组功能的重要方法。对于模式脊椎动物斑马鱼，已为此开发了使用Tol2转座元件的方法。利用这些方法，可以在瞬时和稳定转基因鱼中分析转基因的功能。最近，揭示了Tol2元件转座所需的顺式序列。这使得能够开发出含有易于操作的最小DNA序列的转座子载体。最近，创建并报道了几种含有Gateway序列的转座子载体。这些载体很有用，因为任何外源序列都可以相当容易和快速地克隆到转座子载体中。本章描述了这些转座子载体的特征，以及在斑马鱼中进行转基因的实验方案。