1989
DOI: 10.1128/jvi.63.9.3829-3836.1989
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Cloning and expression of foreign genes in vaccinia virus, using a host range selection system

Abstract: A simple selection system has been developed for the cloning and expression of open reading frames in vaccinia virus. The selection system is based on a conditional lethal (host range) mutant of vaccinia virus. A deletion mutant of the vaccinia virus WR strain was generated by insertion of the neomycin resistance gene from transposon Tn5 and selection with the antibiotic G418. This deletion recombinant, vP293, lacked approximately 21.7 kilobases of DNA beginning 3.8 kilobases from the left end of the genome. v… Show more

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Cited by 146 publications
(28 citation statements)
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“…The vaccinia recombinant expressing mouse p53 cDNA, termed Vp53-wt, was constructed as follows: The DNA fragment encoding mouse full-length wild-type p53 (p53-wt) with a single point mutation in the tetramerization domain was excised from the pGEM-p53 vector (kindly provided by Dr Thomas D. Halazonetis, The Wistar Institute) [36] and cloned into the pSC11 transfer vector for vaccinia virus [37]. A recombinant vaccinia virus containing the p53-wt sequence was generated by homologous recombination with vaccinia virus strain Copenhagen using thymidine kinase-negative cells (TK ± ) [38]. The recombinant viruses were characterized for the presence of the p53 sequence by PCR.…”
Section: Methodsmentioning
confidence: 99%
“…The vaccinia recombinant expressing mouse p53 cDNA, termed Vp53-wt, was constructed as follows: The DNA fragment encoding mouse full-length wild-type p53 (p53-wt) with a single point mutation in the tetramerization domain was excised from the pGEM-p53 vector (kindly provided by Dr Thomas D. Halazonetis, The Wistar Institute) [36] and cloned into the pSC11 transfer vector for vaccinia virus [37]. A recombinant vaccinia virus containing the p53-wt sequence was generated by homologous recombination with vaccinia virus strain Copenhagen using thymidine kinase-negative cells (TK ± ) [38]. The recombinant viruses were characterized for the presence of the p53 sequence by PCR.…”
Section: Methodsmentioning
confidence: 99%
“…11 The M form is under the control of the vaccinia H6 promoter. 12 The L form is under the control of an entomopox early promoter.…”
Section: Canarypox Constructmentioning
confidence: 99%
“…Marker rescue, the principle of the selection system presented here, is well established. In a previous report (Perkus et al, 1989), a host range selection system was described that is based on a parental virus lacking the human host range gene K1L, thus being growth re-FIG. 6.…”
Section: Discussionmentioning
confidence: 99%
“…Restoration of the depleted function by an integration vector allows identification of the recombinant clones. Selection criteria are plaque size (Rodriguez and Esteban, 1989), plaque formation (Blasco and Moss, 1995), and host range (Perkus et al, 1989). In the latter system, the host range gene K1L was deleted from the parental virus, impairing viral growth in certain human cell lines.…”
Section: Introductionmentioning
confidence: 99%