SYNTHETIC CDNA-ENCODING THE RAT AT(1A) RECEPTOR - A USEFUL TOOL FOR STRUCTURE-FUNCTION RELATIONSHIP ANALYSIS

To carry out systematic structure-function studies of the rat angioten sin II receptors by site directed mutagenesis, or production of chimer ic receptors, we have produced a synthetic cDNA coding for the AT1a re ceptor. The synthetic cDNA is 1101 base pairs long, and contains 49 un ique restriction sites that are on the average 23 base pairs apart, al lowing replacement of specific restriction fragments by synthetic coun terparts containing the desired modified sequence. The total cDNA was assembled in the expression vector pECE. After stable expression in Ch inese Hamster Ovary cells, the protein encoded by this synthetic cDNA presents a pharmacological profile and a signal transduction mechanism indistinguishable from the wild type rat AT1a receptor. (C) 1994 Acad emic Press, Inc.