Double digestion: NotI and XbaI yield 7.2 kb and 3.2 kb
Retrovirus can be produced using one of the following methods:
1) Transfection of a retrovirus packaging cell line with a retrovirus expression vector. Packaging cell lines usually stably express gag, pol and env genes. For example, transfection of Plat-E packaging cell line ( MBS169333 ) with a pMXs vector would produce an ecotropic retrovirus.
2) Cotransfection of a host cell with plasmids containing LTRs, Gag, Pol, Env. For example, cotransfection of 293RTV (MBS169332) with pMXs, pCMV-Gag-Pol (MBS168500) and pCMV-VSV-G (MBS168028) would produce VSVG-pseudotyped retrovirus.
Note: We recommend cotransfection of expression vector:gag-pol vector: envelope vector at the following plasmid ratios:
(a) For ecotropic or amphotropic retrovirus, 3:1:1
(b) For VSVG-pseudotyped retrovirus, 3:1:0.5
2. Mann, R., Mulligan, R. C. and Baltimore, D. (1983) Cell 33:153-159.
3. Morita, S., Kojim, T., and Kitamura, T. (2000) Gene Therapy 7: 1063-1066.
Okamoto, K. et al. (2012). Dengue Virus Strain DEN2 16681 Utilizes a Specific Glycochain of Syndecan-2 Proteoglycan as a Receptor. J.Gen. Virol. 93:761-770.
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Product Notes
The pCMV-Gag-Pol (Catalog #AAA168500) is a Retroviral Vector and is intended for research purposes only. The product is available for immediate purchase. It is sometimes possible for the material contained within the vial of "pCMV-Gag-Pol, Retroviral Vector" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
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