Baculovirus AcMNPV Expressing E. coli Cell Line (DE35)
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.
Highlights:
- Cells utilizes the Tn7-based transposition strategy for production of baculovirus genomes in E. coli
- Contains deletions of the viral cathepsin and chitinase genes to help improve protein yield and quality
- The AcMNPV genome is carried on a single-copy F' origin plasmid which can be replicated in E. coli
- Includes protein farnesyltransferase/geranylgeranyltransferase type-1 subunit alpha (FNTA) and protein farnesyltransferase subunit beta (FNTB)
- See also: KRAS4b Baculovirus Expression Vectors
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.
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.
Highlights:
- Cells utilizes the Tn7-based transposition strategy for production of baculovirus genomes in E. coli
- Contains deletions of the viral cathepsin and chitinase genes to help improve protein yield and quality
- The AcMNPV genome is carried on a single-copy F' origin plasmid which can be replicated in E. coli
- Includes protein farnesyltransferase/geranylgeranyltransferase type-1 subunit alpha (FNTA) and protein farnesyltransferase subunit beta (FNTB)
- See also: KRAS4b Baculovirus Expression Vectors
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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| Product Type: | Cell Line |
| Name: | DE35 |
| Cell Type: | Bacterial |
| Organism: | E. coli |
| Biosafety Level: | BSL-1 |
| Growth Conditions: | LB or other rich media, 30 minute doubling at 37C |
| Cryopreservation: | 25% glycerol |
| Comments: | 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. |
| Storage: | -80C |
| Shipped: | Dry Ice |
- Gillette WK, Esposito D, Abreu Blanco M, Alexander P, Bindu L, Bittner C, Chertov O, Frank PH, Grose C, Jones JE, Meng Z, Perkins S, Van Q, Ghirlando R, Fivash M, Nissley DV, McCormick F, Holderfield M, Stephen AG. Farnesylated and methylated KRAS4b: high yield production of protein suitable for biophysical studies of prenylated protein-lipid interactions. Sci Rep. 2015 Nov 2;5:15916. doi: 10.1038/srep15916. PubMed PMID: 26522388; PubMed Central PMCID: PMC4629113.
- Gillette W, Frank P, Perkins S, Drew M, Grose C, Esposito D. Production of Farnesylated and Methylated Proteins in an Engineered Insect Cell System. Methods Mol Biol. 2019;2009:259-277. View Article
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