EFFICIENT PRODUCTION OF HUMAN GAMMA-INTERFERON IN TOBACCO PROTOPLASTSBY GENETICALLY-ENGINEERED BROME MOSAIC-VIRUS RNAS

Citation
M. Mori et al., EFFICIENT PRODUCTION OF HUMAN GAMMA-INTERFERON IN TOBACCO PROTOPLASTSBY GENETICALLY-ENGINEERED BROME MOSAIC-VIRUS RNAS, Journal of General Virology, 74, 1993, pp. 1255-1260
Citations number
22
Categorie Soggetti
Virology
Journal title
ISSN journal
00221317
Volume
74
Year of publication
1993
Part
7
Pages
1255 - 1260
Database
ISI
SICI code
0022-1317(1993)74:<1255:EPOHGI>2.0.ZU;2-5
Abstract
We succeeded in producing human gamma interferon (IFN-gamma) in tobacc o protoplasts in quantity using genetically engineered brome mosaic vi rus (BMV strain ATCC66). This strain of BMV produces two types of coat protein, a full-length coat protein (20K) and a truncated coat protei n (19K) which are translated from the first and second initiation codo ns, respectively. We replaced the truncated coat protein gene with the IFN-gamma gene and synthesized BMV-IFN-gamma chimera RNAs using an in vitro transcription system. The BMV-IFN-gamma chimera RNAs were used to inoculate tobacco protoplasts together with BMV RNA 1 and RNA 2 and produced IFN-gamma to a level of 5 to 10 % of total extracted protein s per infected protoplast after 24 h of incubation. The efficient prod uction of IFN-gamma was attributed to the high translation activity of the BMV-IFN-gamma chimera RNA. We demonstrate that 24 nucleotides cod ing for the N-terminal amino acids of the full-length coat protein wer e probably involved in the high translation activity of the BMV-IFN-ga mma chimera RNA.