RRC ID 209
Author Koga A, Hori H, Sakaizumi M.
Title Gene transfer and cloning of flanking chromosomal regions using the medaka fish Tol2 transposable element.
Journal Mar Biotechnol (NY)
Abstract For the ultimate purpose of developing genetic tools using the medaka fish Tol2 transposable element, we examined whether it can transfer a marker gene into the fish genome and also be applied for cloning of chromosomal regions adjacent to insertion points. An internal region of Tol2 was removed and replaced with the green fluorescent protein (GFP) gene and a bacterial plasmid replication origin. This modified Tol2 clone was microinjected into fertilized eggs together with messenger RNA for the Tol2 transposase. The GFP gene was found to be integrated into chromosomes and transmitted to subsequent generations. Restriction enzyme digestion of genomic DNA of a transformant fish, followed by ligation and introduction into bacteria, produced a plasmid containing the entire element and flanking chromosomal regions. Sequencing analysis of this clone demonstrated transposition of the element in the germline of the first generation. Thus, the basic requirements for a gene transfer vector and gene tagging system were fulfilled.
Volume 4(1)
Pages 6-11
Published 2002-1-1
DOI 10.1007/s10126-001-0086-2
PMID 14961283
IF 2.798
Times Cited 10
Medaka O. luzonensis (RS270)