CLONING, STRUCTURAL ORGANIZATION, AND TRANSCRIPTIONAL ACTIVITY OF THERAT VITAMIN-K-DEPENDENT GAMMA-GLUTAMYL CARBOXYLASE GENE

The vitamin K-dependent gamma-glutamyl carboxylase gene was cloned fro m a rat liver genomic DNA library and the structural organization of t his gene was determined. The carboxylase gene is 16.3 kb in length and contains 15 exons and 14 introns. DNA sequence analysis revealed that all 14 introns were U2-Type GT-AG introns. A 2.8-kb DNA fragment corr esponding to the 5'-flanking region of the cloned gene demonstrated tr anscriptional activity in a rat liver cell. line that is known to expr ess the endogenous carboxylase gene. DNA sequence analysis of the prox imal 331 bp of this 5'-flanking sequence reveals the absence of an ide ntifiable TATA box. Consensus sequences for several transcription fact ors that may be important in regulating its tissue specific expression were identified. The isolation and characterization of the rat carbox ylase gene provides essential information for the analysis of its regu lation in vitro and in transgenic animal models. (C) 1998 Academic Pre ss.