Utilization of vaccinia virus promoters by fowlpox virus recombinants

Schnitzlein, W.M.; Tripathy, D.N.

Animal Biotechnology 1(2): 161-174

1990


ISSN/ISBN: 1049-5398
DOI: 10.1080/10495399009525738
Accession: 002272778

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Abstract
The ability of fowlpox virus (FPV) to recognize vaccinia virus promoters was indicated by the transient expression of .beta.-galactosidase (.beta.-gal) when a plasmid containing a vaccinia P11 promoter-bacterial lacZ gene fusion was transfected into FPV-infected cells. Based on this apparent utilization of a heterologous poxvirus promoter, FPV recombinants capable of expressing .beta.-gal were created. For this purpose, plasmids which would direct the insertion of a foreign transcriptional unit into the FPV thymidine kinase (TK) gene were constructed. Following plasmid transfection of FPV-infected monolayers, the recombinants in the progeny were partially selected due to their TK- phenotype and visually identified by screening plaques for the presence of .beta.-gal. The predicted location of the foreign DNA in the recombinant FPV genome was verified by restriction enzyme analysis and subsequent hybridization studies. A second recombinant, in which the vaccinia late P11 promoter had been replaced by the early-late P7.5 promoter, was also created. The transcriptional regulatory natures of both translocated promoters were retained, even though they were less efficient in the heterologous virus.