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Expression cloning of membrane proteins in Xenopus oocytes
Department of Physiology and Pharmacology, University of Queensland
Fig. 1. Expression cloning strategy in _Xenopus_ oocytes. The right panel shows the different cloning steps graphically. The left panel represents the transport assay with the expected results at the corresponding stage of the project. The strategy in brief: 1) Isolation of total poly (A)+ RNA from rabbit renal cortex followed by injection and transport assay (I) in oocytes. 2) Fractionation of the mRNA, injection of the fractions and transport assay (II). 3) Construction of a cDNA library from rabbit renal cortex mRNA. Division of the library into different pools of clones. Synthesis of in vitro transcribed mRNA, injection and transport assay (III). 4) Subdivision of the pool(s) as described above down to one single clone (IV).