A simplified method for the production of recombinant baculovirus.
A simplified method for producing recombinant baculovirus for expression of foreign genes is described. The method utilizes insect cells infected with the wild-type virus before transfection with the plasmid transfer vector, instead of the standard procedure utilizing cotransfection with a plasmid and viral DNA. Recombinant virus is preselected by a limiting dilution dot-blot hybridization procedure, rather than by morphologic criteria alone. In addition, we have found that plasmid purification by anion-exchange chromatography is as efficacious for transfection as plasmid purified by cesium chloride density gradient centrifugation. These modifications allows for an efficient, rapid, inexpensive and more objective protocol for the selection of recombinant baculovirus compared to the conventional protocol.
Goswami
B B
BB
Georgetown University School of Medicine.
Glazer
R I
RI
D000595:Amino Acid Sequence; D000818:Animals; D016367:Baculoviridae / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D015139:Blotting, Southern; D015153:Blotting, Western; D002852:Chromatography, Ion Exchange; D003001:Cloning, Molecular / Q000379:methods*; D005822:Genetic Vectors; D015151:Immunoblotting / Q000379:methods; D007527:Isoenzymes; D008969:Molecular Sequence Data; D010957:Plasmids; D011493:Protein Kinase C / Q000096:biosynthesis / Q000235:genetics; D014162:Transfection
D007527:Isoenzymes; D011493:Protein Kinase C