pHippy plasmid vector质粒载体 BioVector NTCC质粒载体菌种细胞基因保藏中心

pHippy plasmid vector质粒载体

The PhoenixTM Retrovirus Expression Kit includes: an Ecotropic, an Amphotropic
packaging cell line, and two retroviral expression vectors for high-level protein expression in
most mammalian cells.
Contents:
106 PhoenixTM ampho packaging cells
106 PhoenixTM eco packaging cells
10 µg pBMNZ plasmid
10 µg pBMN-GFP plasmid
PhoenixTM Retroviral Expression System Introduction
The Phoenix™ retroviral system is currently used in over 2500 laboratories worldwide for delivery of genes and libraries to cells for biomedical research. An Ecotropic packaging
system or an Amphotropic system is currently available for the delivery of genes to most
dividing mammalian cell types.
Phoenix™ helper-free retrovirus producer lines:
PhoenixTM retrovirus producer lines, Phoenix™-Eco and Phoenix™-Ampho (reference),
for the generation of helper free ecotropic and amphotropic retroviruses are based on the 293T
cell line (a human embryonic kidney line transformed with adenovirus E1a and carrying a
temperature sensitive T antigen co-selected with neomycin), which is highly transfectable
using either calcium phosphate mediated transfection (see section VI) or lipid-transfection
protocols.
These retrovirus producer lines were created by placing into 293T cells constructs capable of
producing gag-pol and envelope protein for ecotropic and amphotropic viruses. The lines
offer many advantages over previous stable systems and these have led to their broad
application worldwide.
1. Virus can be produced in just a few days.
2. By introducing an IRES-CD8 surface marker downstream of the reading frame of the gagpol construct, CD8 expression is a direct reflection of intracellular gag-pol and the stability of
the producer cell population's ability to produce gag-pol can be readily monitored by flow
cytometry.
3. To prevent potential recombination, different non-Moloney promoters for gag-pol and
envelope were used.
4. Both lines are capable of carrying episomes for long-term stable retrovirus production.
5. Helper virus production in these lines have been extensively tested and found to be helpervirus free.
Phoenix™ retrovirus vectors
Encoding either LacZ or GFP, these Moloney murine leukemia virus based vectors were
specifically designed to facilitate the optimalization of protein expression in a variety of
mammalian cells utilizing X-gal staining or FACS analysis. For comprehensive reviews of
retrovirus biology, please refer to listed publications (Section V).
pBMN-GFP
Catalog No: PVL-10014
Quantity: 10 µg in 10 µl of TE (10 mM Tris HCl pH 7.5, 1mM EDTA)
This EBV/retrovirus hybrid plasmid carries gene encoding green fluorescent protein (GFP),
internal ribosomal entry site (IRES), and the Epstein Bar virus nuclear antigen (EBNA) gene
that allows the establishment of stable episomes at 5-20 copies per cell.
Reference: Heemskerk, M. H. M., Blom, B., Nolan, G., Stegmann, A. P. A., Bakker, A. Q.,
Weijer, K., Res, P. C. M. & Spits, H. (1997) J. Exp. Med. 186, 1597-1602.
pBMN-Z
Catalog No: PVL-10011
Quantity: 10 µg in 10 µl of TE (10 mM Tris HCl pH 7.5, 1mM EDTA)
This parental retroviral vector expresses the gene of interest under the control of viral LTR
and carries Ampicillin resistance.

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