Abstract
Plasmids were constructed fusing vaccinia transcriptional regulatory sequences (promoters) to the lacZ gene of Escherichia coli. These recombinant plasmids were used to compare relative promoter strengths in transient expression assays and to construct recombinant vaccinia viruses producing β-galactosidase (βGal). Viruses synthesizing βGal were determined by utilizing the chromogenic substrate, 5-bromo-4-chloro-3-indoyl-β- d-galactoside to form blue plaques. A recombinant virus producing βGal was then used to select a second recombinant virus. This was accomplished via in vivo recombination replacing the lacZ gene with a sequence coding for the gp85 protein of Friend murine leukemia virus. The recombinant virus was selected by its inability to form blue plaques under appropriate conditions.
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