STUDIES ON TECHNIQUES FOR INTRACELLULAR INTRODUCTION OF MACROMOLECULES

Publisher Summary This chapter discusses the use of the silkworm Bombyx mori nuclear polyhedrosis virus (BmNPV) system for the hyperproduction of ts-src, v-myc, v-erbB, and v-sis products as a fusion protein with the viral protein polyhedrin (NP). Fluoresceinisothiocyanate (FITC)-bovine serum albumin (BSA) and FITC-dextran are used as markers, and these markers are introduced into L cells in high frequency with improved liposome techniques. Chimeric oncogenes with the polyhedrin gene of BmNPV can be highly expressed as fused proteins. In case of v-src and v-erbB genes, their products show tyrosine kinase activity even in a fused form. Purification of the products seems to be a little bit difficult as they are insoluble in water. Cleavage of the domain of the oncogene from a fused protein and its purification without loss of biological function remains to be solved. The chapter describes the effort to improve the liposome methods and presents the development of new methods with which constant and high efficiency introduction can be obtained.