A new vector for efficient generation of p10-single-late-promoter recombinant baculoviruses.
A new baculovirus (BacTen) was constructed in order to generate p10 recombinant expression vectors at high frequency. This virus is an AcMNPV derivative, with the polyhedrin gene deleted and thus exhibiting p10 promoter as a single strong late promoter. The polyhedrin coding sequence was re-inserted subsequently under the control of the p10 promoter, in place of the p10 coding sequence. Two flanking Bsu36I restriction sites were inserted together with the polyhedrin coding region. BacTen can, therefore, be efficiently restricted at the p10 locus and used in co-transfection experiments along with p10 transfer vectors carrying the foreign gene to be expressed. It is shown with three independent transfer vectors, that the proportion of recombinants in the viral progeny can be as high as 80% The BacTen baculovirus represents a new powerful tool for the generation of p10 promoter based expression vectors. in a system without the background of considerable production of very late proteins.
Chaabihi
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Proteine Performance S.A., Station de Recherches, Saint Christol Lez Alès, France.
Cêtre
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Berne
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D000818:Animals; D002460:Cell Line; D004279:DNA, Viral; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism; D005822:Genetic Vectors; D017924:Nucleopolyhedroviruses / Q000235:genetics*; D011401:Promoter Regions, Genetic; D011995:Recombination, Genetic; D018411:Spodoptera / Q000166:cytology; D014764:Viral Proteins / Q000235:genetics*
D004279:DNA, Viral; D014764:Viral Proteins; C078072:p10 protein, Baculovirus; D015252:Deoxyribonucleases, Type II Site-Specific; C062194:endodeoxyribonuclease SauI