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Abstract for pSV-β-Galactosidase Control Vector
The pSV-β-Galactosidase Control Vector is designed as a positive control
vector for monitoring transfection efficiencies of mammalian cells. The SV40
early promoter and enhancer drive transcription of the bacterial lacZ
gene, which in turn, is translated into the β-galactosidase enzyme. β-galactosidase
is an excellent reporter enzyme that can be assayed quickly and directly in cell
extracts using spectrophotometric, fluorescent or chemiluminescent assays. This
reporter enzyme is also widely used for in situ histochemical analysis using the
substrate X-Gal.
The pSV-β-Galactosidase Control Vector can be co-transfected with your DNA
of interest. For example, co-transfection with firefly luciferase gene vectors
(pGL3 Vectors) provide cell extracts that can be assayed for both luciferase and
β-galactosidase activities. In this manner, the pSV-β-Galactosidase Vector
acts as an internal control for transient expression assays. A negative control
extract, prepared from mock-transfected cells, should also be assayed for the
presence of endogenous β-galactosidase activity in cultured cells. In
addition, co-transfection with chloramphenicol acetyltransferse reporter gene
vectors (pCAT®3 Vectors) permits assaying for both CAT and β-galactosidase
activities.
The pSV-β-Galactosidase Vector is a modification of pRSV-βGAL with SV40
and pUC18 sequences substituted for RSV and pBR322 sequences. The pSV-β-Galactosidase Vector will express
β-galactosidase in E. coli
due to the presence of the E. coli gpt promoter located upstream of the
lacZ gene. Colonies of E. coli containing the pSV-β-Galactosidase Vector will appear blue when plated on media containing
X-gal. |
