Rapid purification of recombinant baculovirus using fluorescence-activated cell sorting.
Expression of foreign proteins in the baculovirus-insect cell expression system has been limited by difficulties in rapid identification and purification of recombinant virus. Although the process of identifying recombinant virus has been greatly facilitated by the introduction of vectors that lead to insect cell co-expression of beta-galactosidase with foreign genes of interest, isolation of pure recombinant virus using plaque purification may still take several weeks to months to accomplish. Using a fluorescent beta-galactosidase substrate, we have established that insect cells harboring recombinant virus can be rapidly isolated using fluorescence-activated cell sorting. Pure recombinant virus can then be readily obtained using this cellular fraction, with a pure viral culture generally obtained within 2-3 weeks of insect cell transfection.
Peng
S
S
Department of Physiology and Biophysics University of Alabama, Birmingham 35294-0005.
Sommerfelt
M A
MA
Berta
G
G
Berry
A K
AK
Kirk
K L
KL
Hunter
E
E
Sorscher
E J
EJ
D000818:Animals; D016367:Baculoviridae / Q000235:genetics / Q000302:isolation & purification*; D002469:Cell Separation; D002478:Cells, Cultured; D004274:DNA, Recombinant; D005434:Flow Cytometry; D005822:Genetic Vectors; D009036:Moths; D014162:Transfection; D001616:beta-Galactosidase / Q000235:genetics
D004274:DNA, Recombinant; D001616:beta-Galactosidase