E. coli DE35 cells with the complete Autographa californica nucleopolyhedrovirus (AcMNPV) baculovirus genome which has been modified to produce enzymes, required for high yield farnesylation in insect cells.
DE35 is an analogous strain to E. coli DH10Bac cells, which utilizes the Tn7-based transposition strategy for production of baculovirus genomes in E. coli. The modified AcMNPV genome is carried on a single-copy F’ origin plasmid which can be replicated in E. coli and a subsequent alkaline lysis DNA preparation can be used to transfect insect cells to produce competent baculovirus. In addition to the introduction of human prenylation-related enzymes FNTA and FNTB , the viral genome in DE35 also includes deletions of the viral cathepsin and chitinase genes to improve protein yield and quality.
From the laboratory of Dominic Esposito, PhD, National Cancer Institute/NIH.
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|LB or other rich media, 30 minute doubling at 37C
|Glycerol stocks of E. coli DE35 cells should be propagated with 50 ug/ml kanamycin and 7 ug/ml tetracycline to ensure bacmid and helper plasmid stability to maintain plasmids.
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