A new method for the isolation of recombinant baculovirus.
An improved method for the isolation of baculovirus recombinants is described. The method involves the replication and maintenance of the baculovirus genome in the yeast Saccharomyces cerevisiae which was accomplished by the isolation of a baculovirus recombinant containing yeast ARS and CEN sequences ensuring stable replication in yeast and a URA3 selectable marker. The viral DNA maintained its ability to replicate in insect cells. An efficient and rapid selection system was set up, to isolate viral recombinants in yeast; DNA from selected yeast colonies was transfected into insect cells to obtain recombinant virus. We demonstrate the utility of this system by isolating recombinant viruses that express two different members of the CREB/ATF family of transcription factors.
Patel
G
G
Gene Regulation Laboratory, Imperial Cancer Research Fund, Lincoln's Inn Fields, London, UK.
Nasmyth
K
K
Jones
N
N
D000818:Animals; D016367:Baculoviridae / Q000235:genetics* / Q000302:isolation & purification; D001483:Base Sequence; D002460:Cell Line; D017362:Cyclic AMP Response Element-Binding Protein; D004274:DNA, Recombinant / Q000235:genetics*; D004279:DNA, Viral / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000235:genetics*; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D007313:Insecta / Q000235:genetics; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics; D014157:Transcription Factors / Q000235:genetics*
D017362:Cyclic AMP Response Element-Binding Protein; D004274:DNA, Recombinant; D004279:DNA, Viral; D004268:DNA-Binding Proteins; D014157:Transcription Factors