pCMVBeta Mammalian lacZnls12co Expression Vector

Product ID: M1017



Unit SizePriceQuantity 
20ug
$336.96

Availability: In stock


Description

High copy number eukaryotic vector, pCMVβ expresses the full-length codon-optimized β-galactosidase gene (lacZco) with an effective nuclear localization signal under the control of the cytomegalovirus immediate early gene (CMV IE) promoter. The nls12 variant results from addition of a twelve amino acid sequence from SV40 T antigen nuclear location signal after the methionine initiation residue.

When expressed in mammalian cells, the codon-optimized gene results in expression levels of β-galactosidase 15-fold higher than those resulting from an analogous construct containing the native E. coli gene sequence. Enhanced transcript stability and increased translational efficiency provide for increased β-galactosidase expression, as suggested by RNA analysis. In addition, codon-optimization results in the elimination of several cryptic splice acceptor sites that are present in the native E. coli gene sequence and increases the amounts of un-spliced, full-length genomic RNA when used in a lentiviral vector containing a 5' splice donor. The β-galactosidase enzyme expression is also enhanced by the SV40 late polyadenylylation signal. This pCMVβ expression vector contains the β-lactamase gene, which acts as a selection marker (100μg/mL ampicillin resistance) in E. coli host. pCMVβ vector has been tested to generate up to 2530u/mg cell extract (MacGregor, and Caskey). If desired, the β-galactosidase codon optimized gene can be excised using XhoI and NotI sites to allow the insertion of other genes to be expressed under the same regulatory elements in mammalian cells. See also our pSV40β lacZnls12co vector system, M1018.

Technical Data
SKU M1017
Unit Size 20ug
Notes

Full Length Sequence Data

Restriction Sites


References and Citations

Citations:

  • Jubeli E, Maginty AB, Abdul Khalique N, Raju L, Abdulhai M, Nicholson DG, Larsen H, Pungente MD, Goldring WP. (2015) "Next generation macrocyclic and acyclic cationic lipids for gene transfer: Synthesis and in vitro evaluation." Bioorg Med Chem. 23(19):6364-78
  • Øpstad CL, Zeeshan M, Zaidi A, Sliwka HR, Partali V, Nicholson DG, Surve C, Izower MA, Bilchuk N, Lou HH, Leopold PL, Larsen H, Liberska A, Khalique NA, Raju L, Flinterman M, Jubeli E, Pungente MD. (2014) "Novel cationic polyene glycol phospholipids as DNA transfer reagents--lack of a structure-activity relationship due to uncontrolled self-assembling processes." Chem Phys Lipids. 183:117-36.
  • Parvizi P, Jubeli E, Raju L, Khalique NA, Almeer A, Allam H, Manaa MA, Larsen H, Nicholson D, Pungente MD, Fyles TM. (2014) "Aspects of nonviral gene therapy: correlation of molecular parameters with lipoplex structure and transfection efficacy in pyridinium-based cationic lipids." Int J Pharm. 461(1-2):145-56.
  • Øpstad CL, Sliwka HR, Partali V, Elgsaeter A, Leopold P, Jubeli E, Khalique NA, Raju L, Pungente MD. (2013) "Synthesis, self-assembling and gene delivery potential of a novel highly unsaturated, conjugated cationic phospholipid." Chem Phys Lipids. 170-171:65-73.
  • Yeh J, Tamarkina E, Khalique NA, Raju L, Pungente MD. (2012). "Synthesis, self-assembly and lipoplex formulation of two novel cyclic phosphonate lipids." QScience Connect: Vol. 2012, 6.
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References:

  • Anson DS, Limberis M. (2004). "An improved β-galactosidase reporter gene." J. Biotech. 108: 17-30.
  • MacGregor GR, Caskey CT. (1989). "Construction of plasmids that express E. Coli beta-galactosidase in mammalian cells." Nucleic Acids Res. 17: 2365.
  • Norton PA, Coffin JM. (1984). "Bacterial b-galactosidase as a marker of rous sarcoma virus gene sxpression and replication." Mol. Cell Biol. 5: 281.
  • Nolan GP. (1988) "Fluorescence-activated cell analysis and sorting of viable mammalian cells based on b-D-galactosidase activity after transduction of Escherichia coli lacZ." Proc. Natl. Acad. Sci (USA). 85: 2603.
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