Molecular Cloning, Encoding Sequence, and Expression of Vaccinia Virus Nucleic Acid-Dependent Nucleoside Triphosphatase Gene
A rabbit poxvirus genomic library contained within the expression vector λ gt11 was screened with polyclonal antiserum prepared against vaccinia virus nucleic aciddependent nucleoside triphosphatase (NTPase)-I enzyme. Five positive phage clones containing from 0.72- to 2.5-kilobase-pair (kbp) insert...
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Veröffentlicht in: | Proceedings of the National Academy of Sciences - PNAS 1986-12, Vol.83 (24), p.9566-9570 |
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Sprache: | eng |
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Zusammenfassung: | A rabbit poxvirus genomic library contained within the expression vector λ gt11 was screened with polyclonal antiserum prepared against vaccinia virus nucleic aciddependent nucleoside triphosphatase (NTPase)-I enzyme. Five positive phage clones containing from 0.72- to 2.5-kilobase-pair (kbp) inserts expressed a β -galactosidase fusion protein that was reactive by immunoblotting with the NTPase-I antibody. Hybridization analysis allowed the location of this gene within the vaccinia HindIIID restriction fragment. From the known nucleotide sequence of the 16-kbp vaccinia HindIIID fragment, we identified a region that contains a 1896-base open reading frame coding for a 631-amino acid protein. Analysis of the complete sequence revealed a highly basic protein, with hydrophilic COOH and NH2 termini, various hydrophobic domains, and no significant homology to other known proteins. Translational studies demonstrate that NTPase-I belongs to a late class of viral genes. This protein is highly conserved among Orthopoxviruses. |
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ISSN: | 0027-8424 1091-6490 |
DOI: | 10.1073/pnas.83.24.9566 |