A new, rapid and simple procedure for direct cloning of PCR products into baculoviruses.
We propose a novel method for direct cloning of foreign genes into baculoviruses which avoids the use of bacterial transfer vectors. The foreign gene to be inserted is derived by PCR using appropriate primers each of which contains an additional 50 nt of baculovirus sequence for homologous recombination between the PCR-derived DNA and the baculovirus DNA, thus accomplishing insertion of the foreign gene into the baculovirus. The direct cloning of green fluorescent protein and beta-glucuronidase in different baculovirus loci is described. The method is simple and avoids the use of cumbersome techniques associated with enzymatic treatment and DNA purification.
Gritsun
T S
TS
Institute of Virology and Environmental Microbiology, Mansfield Road, Oxford OX1 3SR, UK.
Mikhailov
M V
MV
Roy
P
P
Gould
E A
EA
D016367:Baculoviridae / Q000235:genetics*; D001483:Base Sequence; D003001:Cloning, Molecular / Q000379:methods*; D017931:DNA Primers; D004279:DNA, Viral; D005966:Glucuronidase / Q000235:genetics; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000235:genetics; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction
D017931:DNA Primers; D004279:DNA, Viral; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins; D005966:Glucuronidase