A gene trap approach in Xenopus.

Bronchain, O.J., Hartley, K.O. and Amaya, E.

Current Biology, 9:1195-1198 (1999) (Full Article, 120K)

Abstract

The frog transgenesis technique ultimately promises to make mutagenesis possible through random

insertion of plasmid DNA into the genome. This study was undertaken to evaluate whether a gene trap

approach combined with transgenesis would be appropriate for performing insertional mutagenesis in

Xenopus embryos. Firstly, we confirmed that the transgenic technique results in stable integration into the

genome and that transmission through the germline occurs in the expected Mendelian fashion. Secondly,

we developed several gene trap vectors, using the green fluorescent protein (GFP) as a marker. Using

these vectors, we trapped several genes in Xenopus laevis that are expressed in a spatially restricted

manner, including expression in the epiphysis, the olfactory bulb and placodes, the eyes, ear, brain,

muscles, tail and intestine. Finally, we cloned one of the trapped genes using 5' rapid amplification of

cDNA ends polymerase chain reaction (RACE PCR). These results suggest that the transgenic technique

combined with a gene trap approach might provide a powerful method for generating mutations in

endogenous genes in Xenopus.

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