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pBMN-i-EGFP vector retroviral vector逆转录病毒荧光表达载体质粒 BioVector NTCC质粒载体菌种细胞基因保藏中心

  • 价  格:¥39865
  • 货  号:pBMN-i-EGFP vector
  • 产  地:北京
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pBMN-i-EGFP vector retroviral vector逆转录病毒荧光表达载体质粒

Retroviral systems
Retroviruses are an efficient means to deliver single DNA expression constructs to a wide range of mammalian cell types. They are by far the easiest and fastest means to deliver genes stably to mammalian cells.
They have an additional advantage in that systems developed in the Nolan Laboratory have allowed for their application to deliver large libraries of genes or peptides (or frankly any DNA-encoded objects) to target cells.
Phoenix-MMULV vectors
Phoenix: The first, termed Phoenix, is currently employed in over 2500 laboratories worldwide for delivery of genes and libraries to cells for biomedical research. It is based on Moloney Murine Leukemia Virus (MMULV) and allows for delivery of genes to most DIVIDING mammalian cell types. The system comes as either an Ecotropic packaging system (capable of delivering genes to dividing murine or rat cells) or an Amphotropic system (capable of delivering genes to dividing cells of most mammalian species, including human). A variant of the system contains simply the gag-pol genes and allows for pseudotyping with alternative envelope proteins such as VSV-G.

Phoenix helper-free retrovirus producer lines:
Phoenix is a second-generation retrovirus producer lines for the generation of helper free ecotropic and amphotropic retroviruses. The lines 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).
The unique feature of this cell line is that it is highly transfectable with either calcium phosphate mediated transfection or lipid-based transfection protocols-- up to 50% or higher of cells can be transiently transfected.
The lines were created by placing into 293T cells constructs capable of producing gag-pol, and envelope protein for ecotropic and amphotropic viruses. The lines offered advantages over previous stable systems in that virus can be produced in just a few days.

The lines have several other improvements over other approaches and this has led to their broad application worldwide:
We added the facility to monitor gag-pol production on a cell-by cell basis by introducing an IRES-CD8 surface marker downstream of the reading frame of the gag-pol construct. Thus, 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.For both the gag-pol and envelope constructs non-moloney promoters were used to minimize recombination potential.Different promoters for gag-pol and envelope were used to minimize their inter-recombination potential.Two cell lines were created, Phoenix-Eco and Phoenix-Ampho. Gag-pol was introduced with hygromycin as the co-selectable marker and the envelope proteins were introduced with diptheria resistance as the co-selectable marker.Finally, a line is available that expresses only gag-pol; this line is termed Phoenix-gp. This line is available for further pseudotyping of retroviral virions with other envelope proteins such as gibbon ape leukemia virus envelope or Vesicular stomatitus VSV-G protein.Both Phoenix-Eco and Phoenix-Ampho have been extensively tested for helper virus production and established as being helper-virus free.Both lines are capable of carrying episomes for long-term stable production of retrovirus (LZRS).
Phoenix Ampho & Eco cell lines - second generation retroviral producer lines (MMULV-based).
PhoenixGP cell line, 293T-derived containing only gag-pol for VSV-G and other pseudotyping.
HEK293T cell line, a human cell line that was derived from the HEK293 cell line by the addition of the SV40 large T antigen. The cell line that can be transfected with plasmid DNA at high efficiency and commonly used for production of retroviral, lentiviral and other viral vectors using the transient transfection method.
pCI-VSVG - VSV-G expression plasmid.
pBMN-LacZ vector - a pBabeMN based retroviral construct with the LacZ gene.pBMN-i-EGFP vector - a pBabeMN based retroviral construct with the EMCV IRES upstream from the humanized/enhanced GFP (EGFP).
pBMN-z-i-neo vector - a pBabeMN based retroviral construct with the EMCV IRES upstream from the Neomycin resistance gene.
Felix vectors (pCPRdEnv; pLionII; pLionIIp-YFP; pTiger and pTiger-YFP) - FIV-based (Feline Immunodeficiency) vectors for delivery to non-dividing cells.

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