期刊名称：Proceedings of the National Academy of Sciences
印刷版ISSN：0027-8424
电子版ISSN：1091-6490
出版年度：1991
卷号：88
期号：4
页码：1511-1515
DOI：10.1073/pnas.88.4.1511
语种：English
出版社：The National Academy of Sciences of the United States of America
摘要：Regulatory elements of the Escherichia coli lac operon were used to construct an inducer-dependent conditional-lethal mutant animal virus. The gene encoding the repressor protein of the lac operon was integrated into the vaccinia virus genome so that it was expressed constitutively, and the lac operator was inserted next to the promoter of a gene that encodes an 11-kDa virion-associated protein of unknown function. The addition of inducer to the cell culture medium provided permissive conditions for isolation of a conditional-lethal mutant virus. Under nonpermissive conditions, the isolated virus did not form plaques, and the yield was decreased by at least 1000-fold under one-step growth conditions. Transcription of the operator-controlled gene was inducer-dependent and necessary for synthesis of the 11-kDa protein. Application of this mutagenesis strategy to other viruses is discussed.
