Chemistry: molecular biology and microbiology – Vector – per se
Patent
1992-09-04
1994-09-20
Schwartz, Richard A.
Chemistry: molecular biology and microbiology
Vector, per se
435 691, 4351723, 4352523, 43525233, 536 231, C12N 1500, C12N 1586, C12N 1511
Patent
active
053488865
ABSTRACT:
A method for producing infectious recombinant baculoviruses in bacteria is described. A novel baculovirus shuttle vector (bacmid) was constructed that contains a low-copy-number bacterial replicon, a selectable drug resistance marker, and a preferred attachment site for a site-specific bacterial transposon, inserted into a nonessential locus of the baculovirus genome. This shuttle vector can replicate in E. coli as a plasmid and is stably inherited and structurally stable after many generations of growth. Bacmid DNA isolated from E. coli is infectious when introduced into susceptible lepidopteran insect cells. DNA segments containing a viral promoter driving expression of a foreign gene in insect cells that are flanked by the left and right ends of the site-specific transposon can transpose to the attachment site in the bacmid propagated in E. coli when transposition functions are provided in trans by a helper plasmid. The foreign gene is expressed when the resulting composite bacmid is introduced into insect cells.
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Luckow, V. A. 1991. Cloning and expression of heterologous genes in insect cells with baculovirus vectors., pp. 97-152. In A. Prokop, R. K. Bajpai, and C. Ho (ed.), Recombinant DNA Technology and Applications. McGraw-Hill, New York.
Lee Stephen C.
Leusch Mark S.
Luckow Verne A.
Olins Peter O.
Bennett Dennis A.
Guzo David
Monsanto Company
Schwartz Richard A.
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