EXPRESSION OF THE VESICULAR STOMATITIS-VIRUS NUCLEOCAPSID PROTEIN GENE IN ESCHERICHIA-COLI - ANALYSIS OF ITS BIOLOGICAL-ACTIVITY INVITRO

The nucleocapsid (N) protein genes of vesicular stomatitis virus, serotypes New Jersey and Indiana, were expressed in Escherichia coli BL21 (DE3) transformed by the recombinant expression vector pET-3a containing N genes under the control of the T7 RNA polymerase promoter. Following induction with isopropylthiogalactopyranoside, the N protein was synthesized, representing 10 to 20% of total cellular protein. The N protein remained in a nonaggregated form in 1 M NaCl solution. The purified N protein was found to be biologically active as demonstrated by its encapsidation of preformed leader RNA as well as its synthesis of genome length RNA in an in vitro transcription-replication reaction. The availability of recombinant N protein in a sufficient amount would facilitate structure and function studies of this important viral protein. © 1993 Academic Press. All rights reserved.