Construction and Preliminary Characterization of a Library of “Lethal” Preterminal Protein Mutant Adenoviruses |
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Authors: | Jerome Schaack William Y Ho Shawna Tolman Elizabeth Ullyat Xiaoling Guo Nina Frank Paul I Freimuth Dick J Roovers and John S Sussenbach |
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Institution: | Department of Microbiology, University of Colorado Health Sciences Center, Denver, Colorado, USA. jerry.schaack@uchsc.edu |
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Abstract: | 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. |
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