The Xenopus Oocyte as an in Vitro Translation and Expression System for Chemosensory — Specific Gene Products
In 1971, Gurdon and coworkers published a seminal paper reporting the first successful in ovo translation of exogenous mRNA into protein. The poly (A+) RNA that was isolated biochemically from red blood cells and microinjected into Xenopus oocytes directed the synthesis of protein globin. Their experiments not only established a system in which to study the in vitro translation of genetic material but also, identified a system in which the functional activity of the translation products could be assayed after expression. In this system, the injected mRNA effectively competes with the endogenous mRNA for the translation machinery to synthesize a variety of proteins. This background synthesis and post-translational modification of functional proteins from endogenous mRNA is particularly advantageous when investigating the functional expression of a multicomponent transduction system that utilizes the G-protein, phospholipase C, inositol triphosphate or the adenylate cyclase 2nd messenger intermediates. More recently, the Xenopus oocyte has been utilized to investigate the expression of neurotransmitter receptors, neuromodulators and voltage-gated ion channels (Dascal, 1987; Snutch, 1988) and the pharmacology of 2nd messengers in the B-adrenergic receptor system (Smith et al., 1987). This technique has also been combined with techniques of recombinant DNA technology to investigate the molecular biology of ion channels (Claudio, 1986), the substance-K receptor (Masu et al., 1987) and visual transduction (Khorana et al., 1988). Recently, several investigators habe been exploring the use of the Xenopus expression system to study chemosensory systems.
KeywordsCitral Alanine Tate Triphosphate Collagenase
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- Colman A (1984) Translation of eukaryotic messenger RNA in Xenopus oocytes. In: Transcription and Translation, a practical approach. BD Hames, SJ Higgins (eds.). IRL Press, Washington, pp 271–300.Google Scholar
- Dascal N (1987) The use of Xenopus oocytes for the study of ion channels CRC Critical Rev Biochem 22:317–387.Google Scholar
- Dascal N, Heldman J, Gershon E, Lancet D (1986) Possible expression of odorant receptor proteins in Xenopus oocytes injected with rat olfactory epithelial mRNA. Soc Neurosci Abstr. 13:369.Google Scholar
- Getchell TV, Grillo M, Tate SS, Urade R, Teeter J, Margolis FL (1989) Expression of catfish amino acid taste receptors in Xenopus oocytes. Science, submitted.Google Scholar
- Margolis FL (1988) Molecular cloning of olfactory-specific gene products. In: Molecular Neurobiology of the Olfactory System. FL Margolis, TV Getchell (eds.) Plenum Press, New York, pp. 237–265.Google Scholar