Product M1018

 

pSV40β lacZnls12co Vector

 

 

Overview:

 

This common eukaryotic expression vector, pSV40b expresses the full-length codon-optimized b-galactosidase gene (lacZco) under the control of simian virus 40 (SV40) early 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.  The nls12 variant results from the addition of a twelve amino acid sequence, ProLysLysLysArgLysValGluAspProLysAsp (from the SV40 T antigen nuclear localization signal) after the methionine initiation residue.1  The b-galactosidase enzyme expression is also enhanced by the SV40 late polyadenylylation signal. The pSV40b expression vector also contains b-lactamase, which acts as a selection marker (100 mg/ml ampicillin resistance) in E. coli host.  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 pCMVb lacZco vector system, Product M1017.

 

Full Length Sequence Data

 

Restriction Sites

 

Price: 20mg  $215.25 (USD) 

 

References:

  1. Anson, D.S., Limberis, M., 2004. “An improved β-galactosidase reporter gene.” J. of Biotech. 108:17-30.
  2. Hall, C.V. et al., 1983. “Expression and Regulation of Escherichia coli lacZ Gene Fusions in Mammalian Cells.” J. Mol. Appl. Gen. 2:101.
  3. Herbomel, P. et al., 1984. “Two distinct enhancers with different cell specificities coexist in the regulatory region of polyoma.” Cell 39:653.
  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.
  5. Nikcevic, G. et al., 2003. “Improved transfection efficiency of cultured human cells.” Cell Biol. Int. 27:735.