Analysis of zebrafish mutants from Ac/Ds transposon screen.

Abstract

Zebrafish is a suitable model organism to research due to certain unique traits which includes; embryo transparency, short generation time, advancement in sequenced genome and high homology percentage. These traits allow zebrafish to be easily studied using mutagenesis. The aim of this project is to screen out homozygous zebrafish mutants generated by Ac/Ds transposon system. The Ds cassette and Ac transposase were co-injected into founder embryos and these fish were raised to adulthood and out-crossed with wild type zebrafish. F1 embryos with florescence reporter gene expression were raised to adulthood and sent for sequencing. Subsequently, zebrafish from each line that were positive for Ds cassettes insertion were in-crossed to generate homozygous mutants. Genomic DNA from progeny fish were analysed by PCR and Southern blot. Homozygous mutants are then characterised for observable phenotypes in early embryonic development. In this project, 45 zebrafish lines have been screened. Carriers of 2 mutant genes, ptenb and SCR, had been found and characterised. Ptenb homozygous mutants were generated for a particular fish line and mutant edema phenotype was observed. However, no conclusive link between the edema phenotype and the ptenb mutant was found. There is a need to screen for more homozygous mutants for characterisation.