The bovine papillomavirus (BPV) is capable of transforming certain rodent fibroblast lines in which the viral DNA remains as a stable extrachromosomal plasmid. These properties have been exploited in developing BPV into a stable extrachromosomal mammalian cell vector. The complete genome cloned into pML2, which is a deletion derivative of pBR322, is capable of serving as a shuttle vector which can replicate as a plasmid in mouse C127 cells or in bacteria. We have studied the expression of the rat preproinsulin gene in BPV vectors in C127 cells and have shown that the expression of the gene is enhancer-dependent. An """"""""enhancer"""""""" element is located in the BPV-1 genome at the 3' end of the transforming region, downstream from the polyadenylation recognition sequence. Using this vector system, a variety of exogenous genes have been expressed. A portion of the human T- cell lymphotropic virus type 1 (HTLV-1) has been expressed off of the mouse metallothionein promoter in a BPV vector. The extrachromosomal state of the DNA should provide a physical characteristic to permit the purification of chromatin complexes of mammalian genes. Using the lac operator, we have developed a technique for rapid purification and identification of sequence- specific binding proteins. This technique, combined with the extrachromosomal papillomavirus plasmid vector system, should facilitate the identification of important viral and cellular gene regulatory proteins. During this year, this project has been phased out and this represents the terminal report on this project.
