Preliminary report on the production of transgenic Oreochromis niloticus using tol 2 kit

Abstract

Pronuclear microinjection of donor plasmid containing the tol 2 transposon that was flanked by a promoter β- actin and green fluorescent protein (GFP) reporter gene were injected into the perinuclear cytoplasm of Oreochromis niloticus fertilized eggs. The aim was to determine if the β-actin promoter will drive the expression of the GFP in specific tissues of the embryo. Few days after hatching, GFP was strongly expressed in the embryos when observed under UV-light microscope. Subsequently, β actin fused with a foreign growth hormone gene in a donor tols 2 transposon along with messenger RNA was co-injected into early stage of fertilized eggs using micro injector. Out of the 30 eggs injected, 15 representing 50% hatchlings were obtained. GH was successfully transposed to the genome of Fo as confirmed from PCR of DNA from fin clip of the founder population. Growth performance in transgenic after 2 months of rearing was 1.06gm (3.7cm) compared with 0.68gm(3.5cm) in nontransgenic individuals. The founder generation is being raised to maturity to determine the level of efficiency in transmission to next generation.

Keywords: Transposon; growth enhancement; Oreochromis niloticus; transgenesis; growth hormone gene