Product M1015

 

pCMVβ lacZco Vector

 

 

Overview:

 

This high copy number eukaryotic vector, pCMVb expresses the full-length codon-optimized b-galactosidase gene (lacZco) under the control of the cytomegalovirus immediate early gene (CMV IE) promoter.  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.1 The b-galactosidase enzyme expression is also enhanced by the SV40 late polyadenylylation signal.  This pCMVb expression vector contains the b-lactamase gene, which acts as a selection marker (100mg/mL ampicillin resistance) in E. coli host.   pCMVb vector has been tested to generate up to 2530u/mg cell extract (MacGregor, and Caskey).  If desired, the b-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 pSV40b lacZco vector system, Product M1016.

 

Full Length Sequence Data

 

Restriction Sites

 

Price: 20mg  $204.75 (USD) 

 

 

References:

  1. Anson, D.S., Limberis, M., 2004. “An improved β-galactosidase reporter gene.” J. of Biotech. 108: 17-30.
  2. MacGregor, G.R. & Caskey, C.T., 1989.  “Construction of Plasmids that Express E. Coli Beta-galactosidase in Mammalian Cells.” Nucleic Acids Res. 17: 2365.
  3. Norton, P.A.& Coffin, J.M., 1984. “Bacterial b-galactosidase as a Marker of Rous Sarcoma Virus Gene Expression and Replication.” Mol. Cell Biol. 5: 281.
  4. Nolan, G.P., et al., 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.