pLVX-MetLuc Control is an HIV-1-based, lentiviral vector that constitutively expresses Metridia luciferase (MetLuc), a secreted reporter protein that can be easily detected in the medium surrounding the cells. The vector can be used as a control to monitor the constitutive expression and secretion of Metridia luciferase in your cell type of interest.
Metridia luciferase is a naturally secreted luciferase from the marine copepod Metridia longa. The human codon-optimized Metridia luciferase gene encodes a 219 amino acid (24 kDa) polypeptide that includes a 17 amino acid N-terminal signal peptide necessary for secretion (1). Expression of Metridia luciferase is driven by the constitutively active human cytomegalovirus immediate early promoter (PCMV IE), located just upstream of the Metridia luciferase coding sequence.
pLVX-MetLuc Control contains all of the viral processing elements necessary for the production of replication-incompetent lentivirus, as well as elements to improve viral titer, transgene expression, and overall vector function. The woodchuck hepatitis virus posttranscriptional regulatory element (WPRE) promotes RNA processing events and enhances nuclear export of viral and transgene RNA (2), leading to increased viral titers from packaging cells, and enhanced expression of your gene of interest in target cells. In addition, the vector includes a Rev-response element (RRE), which further increases viral titers by enhancing the transport of unspliced viral RNA out of the nucleus (3). Finally, pLVX-MetLuc Control also contains a central polypurine tract/central termination sequence element (cPPT/CTS). During target cell infection, this element creates a central DNA flap that increases nuclear import of the viral genome, resulting in improved vector integration and more efficient transduction (4).
In addition to lentiviral elements, pLVX-MetLuc Control contains a puromycin resistance gene (Puror) under the control of the murine phosphoglycerate kinase promoter (PPGK) for the selection of stable transductants. The vector also contains a pUC origin of replication and an E. coli ampicillin resistance gene (Ampr) for propagation and selection in bacteria.
The pLVX-MetLuc Control vector is available as part of the Lenti-X Ready-To-Glow Secreted Luciferase Reporter System (Cat. No. 631746). Lentiviral particles derived from the vector allow you to monitor the constitutive expression and secretion of Metridia luciferase in your cell type of interest. Secretion of Metridia luciferase can be monitored by adding chemiluminescent substrate to a sample of the cell medium. The vector can also be used to monitor the inhibitory effect of drugs, like Brefeldin A, on protein secretion. If required, stable transfectants can be selected using puromycin.
Propagation in E. coli
Recommended host strains: DH5α and other general purpose strains.
Selectable marker: plasmid confers resistance to ampicillin (100 μg/ml) in E. coli hosts.
E. coli replication origin: pUC
References
1. Markova, S.V. et al. (2004) J. Bio. Chem. 279(5):3212-3217.
2. Zufferey, R. et al. (1999) J. Virol. 73(4):2886–2892.
3. Cochrane, A. W. et al. (1990) Proc. Natl. Acad. Sci. USA 87(3):1198–1202.
4. Zennou, V. et al. (2000) Cell 101(2):173–185.
