Expression of the vesicular stomatitis virus nucleocapsid protein gene in Escherichia coli: analysis of its biological activity in vitro.
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.
Department of Molecular Biology, Cleveland Clinic Foundation, Ohio 44195.
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Das
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Banerjee
D001483:Base Sequence; D002213:Capsid / Q000096:biosynthesis* / Q000235:genetics / Q000502:physiology; D004926:Escherichia coli; D015870:Gene Expression / Q000502:physiology; D005814:Genes, Viral / Q000502:physiology*; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000378:metabolism; D012367:RNA, Viral / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis; D014721:Vesicular stomatitis Indiana virus / Q000235:genetics*; D018116:Vesiculovirus; D014758:Viral Core Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000502:physiology
D012333:RNA, Messenger; D012367:RNA, Viral; D011994:Recombinant Proteins; D014758:Viral Core Proteins