Dominant negative selection of vaccinia virus using a thymidine kinase/thymidylate kinase fusion gene and the prodrug azidothymidine
- 作者：Holzer ; Georg W. ; Mayrhofer ; Josef ; Gritschenberger ; Werner ; Falkner ; Falko G.
- 关键词：Dominant selection ; Vaccinia virus ; Thymidine kinase ; Thymidylate kinase ; Viral vectors
- 刊名：Virology
- 出版年：2005
- 期刊代码：108_00426822
- 类别：imm
- 出版时间：July 5, 2005
- 卷：337
- 期：2
- 页码：235-241
- 文件大小：1.80 M
摘要
The Escherichia coli thymidine kinase/thymidylate kinase (tk/tmk) fusion gene encodes an enzyme that efficiently converts the prodrug 3′-azido-2′,3′-dideoxythymidine (AZT) into its toxic triphosphate derivative, a substance which stops DNA chain elongation. Integration of this marker gene into vaccinia virus that normally is not inhibited by AZT allowed the establishment of a powerful selection procedure for recombinant viruses. In contrast to the conventional vaccinia thymidine kinase (tk) selection that is performed in tk-negative cell lines, AZT selection can be performed in normal (tk-positive) cell lines. The technique is especially useful for the generation of replication-deficient vaccinia viruses and may also be used for gene knock-out studies of essential vaccinia genes.