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Viable double vaccinia virus recombinants with the non-inducible phage T7 expression system

Institute of Gene Biology, Russian Academy of Sciences, 117334 Moscow, Vavilov str. 34/5, Russia

Double vaccinia virus recombinants expressing both the T7 RNA polymerase gene, controlled by a weak early poxvirus P_F promoter, and the Escherichia coli -galactosidase gene, controlled by the phage T7 promoter, have been obtained. The viability of the double recombinants depended on the T7 RNA polymerase expression level. If the T7 RNA polymerase gene was inserted into a recombinant already containing the -galactosidase gene, the efficiency of formation of the double recombinants was significantly higher compared to that for the reverse insertion order. The negative effect of the phage T7 terminator on -galactosidase expression in cells infected with the recombinant viruses has been shown. The dynamics and levels of -galactosidase formation by different vaccinia virus recombinants have been studied.

Received 8 July 1992; accepted 15 September 1992.