Abstract
Restriction enzyme mediated integration (REMI) transgenesis and lentiviral transgenesis are effective methods of introducing transgenes into the genome of frogs. One aquatic amphibian species, Xenopus laevis, and one land dwelling species, Litoria caerulea, were chosen as subjects for transgenesis. REMI produced X. laevis that expressed the fluorescent protein DsRed. REMI was unsuccessful in producing transgenic Litoria. Therefore, lentiviral transgenesis was attempted. Hatchling Litoria tadpoles were exposed to replication defective lentiviral particles containing the coding sequence for DsRed. Histological evaluation revealed the presence of DsRed in brain, heart, liver, kidney, and muscle tissues. Therefore, lentiviral transgenesis appears to be a viable technique for producing transgenic land-dwelling frogs. Key words: DsRed, Xenopus laevis, Litoria caerulea, restriction enzyme mediated integration (REMI), Lentivirus.
Highlights
In recent years, the African Clawed Frog Xenopus laevis has become a leading model organism for transgenesis
The development of DsRed transgenic Xenopus would support the use of Xenopus as a model organism because DsRed emits in the visible light spectrum
There was significant difference found between Dulbecco’s Modified Eagle Medium (DMEM) and the other medias, but there is no significant difference between the means of NCTC 109 and Leibovitz 15 (L15) (Statistical Methods Table 4)
Summary
The African Clawed Frog Xenopus laevis has become a leading model organism for transgenesis. Restriction enzyme mediated integration (REMI) of DNA into sperm nuclei, and transplantation into unfertilized eggs, is a method that has been used previously to produce GFP transgenic lines of X. laevis (Sakamaki et al, 2004). Positive results have been produced using the Xenopus cardiac actin promoter, low expression in tissues has been a problem (Sakamaki et al, 2004). The CAG promoter has driven transgenic expression in most Xenopus tissues, including germline cells (Marsh-Armstrong et al, 1999). Transgenic Xenopus progeny show germline transmission without the CAG promoter, but expression is mosaic (Marsh-Armstrong et al., 1999). The development of DsRed transgenic Xenopus would support the use of Xenopus as a model organism because DsRed emits in the visible light spectrum
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