A NEW VECTOR FOR EFFICIENT GENERATION OF P10-SINGLE-LATE-PROMOTER RECOMBINANT BACULOVIRUSES

A new baculovirus (BacTen) was constructed in order to generate p10 re combinant expression vectors al high frequency. This virus is an AcMNP V derivative, with the polyhedrin gene deleted and thus exhibiting p10 promoter as a single strong late promoter. The polyhedrin coding sequ ence was re-inserted subsequently under the control of the p10 promote r, in place of the p10 coding sequence. Two flanking Bsu36I restrictio n sites were inserted together with the polyhedrin coding region. BacT en can, therefore, be efficiently restricted at the p10 locus and used in co-transfection experiments along with p10 transfer vectors carryi ng the foreign gene to be expressed. It is shown with three independen t transfer vectors, that the proportion of recombinants in the viral p rogeny can be as high as 80%. The BacTen baculovirus represents a new powerful tool for the generation of p10 promoter based expression vect ors, in a system without the background of considerable production of very late proteins. Copyright (C) 1997 Elsevier Science B.V.