Common eukaryotic expression vector, pSV40β expresses the full-length codon-optimized β-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 β-galactosidase enzyme expression is also enhanced by the SV40 late polyadenylylation signal. The pSV40β expression vector also contains β-lactamase, which acts as a selection marker (100 μg/ml ampicillin resistance) in E. coli host. 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 pCMVβ lacZco vector system, M1015.
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