Construction and preliminary characterization of a library of "lethal" preterminal protein mutant adenoviruses.
Adenoviruses containing lethal in-frame insertion mutant alleles of the preterminal protein (pTP) gene were constructed with cell lines that express pTP. Thirty in-frame insertion mutant alleles, including 26 alleles previously characterized as lethal and 4 newly constructed mutant alleles, were introduced into the viral chromosome in place of the wild-type pTP gene. The viruses were tested for ability to form plaques at 37 degrees C in HeLa-pTP cells and at 32 degrees C and 39.5 degrees C in HeLa cells. Two of the newly constructed viruses exhibited temperature sensitivity for plaque formation, one virus did not form plaques in the absence of complementation, seven additional mutants exhibited a greater than 10-fold reduction in plaque formation in the absence of complementation, and another eight mutants exhibited stronger phenotypes than did previously characterized in-frame insertion mutants in the plaque assay. These mutant viruses offer promise for analysis of pTP functions.
Schaack
J
J
Department of Microbiology, University of Colorado Health Sciences Center, Denver, Colorado, USA. jerry.schaack@uchsc.edu
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Tolman
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Ullyat
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Guo
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Frank
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Freimuth
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Roovers
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Sussenbach
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D000260:Adenoviruses, Human / Q000235:genetics*; D015723:Gene Library; D005804:Genes, Lethal; D006801:Humans; D016254:Mutagenesis, Insertional; D010750:Phosphoproteins / Q000235:genetics*; D011498:Protein Precursors / Q000235:genetics*; D014407:Tumor Cells, Cultured; D014764:Viral Proteins / Q000235:genetics
D010750:Phosphoproteins; D011498:Protein Precursors; D014764:Viral Proteins; C085476:pTP protein, adenovirus