Quantitative assessment of poxvirus promoters in fowlpox and vaccinia virus recombinants

Abstract

A comparison was undertaken of poxvirus promoters in vaccinia and fowlpox virus (FPV) recombinants using the level of beta-galactosidase expressed from the LacZ gene as a measure of promoter function. In this study a comparison was made of the vaccinia virus promoters, P 7.5 and P L11, the major late promoter of cowpox virus, P CPX (expressing the abundant inclusion body protein), and the FPV promoters, P E/L and P L. In vaccinia virus recombinants the FPV P E/L promoter expressed one-third to one-half the level of beta-galactosidase expressed by the P L11 promoter. In comparison with the P 7.5 promoter, the FPV P E/L promoter expressed four to five times the level of beta-galactosidase. In FPV recombinants beta-galactosidase activity expressed was equal for the P E/L and P CPX promoters. Levels expressed by P L11 and P L were one-half and one-fifth that level, respectively. The temporal regulation of the promoters was maintained in both vaccinia virus and FPV recombinants. The P E/L promoter of FPV has the TAAATG sequence characteristic of late poxvirus promoters at the transcription initiation site. In an attempt to enhance the utility of this promoter for the expression of foreign genes in FPV and vaccinia virus recombinants, the effect upon promoter function of changing the G of the ATG to A, T, or C was determined using transient expression assays with vaccinia virus. Substitution of A, T, or C for the G abolished promoter function. Because of its early/late function, the level of expression and the presence of the oppositely oriented late P L promoter, the FPV P E/L promoter will be valuable for the expression of foreign genes in poxvirus recombinants.