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Journal of Virology, November 1999, p. 9599-9603, Vol. 73, No. 11
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Construction and Preliminary Characterization of a Library of "Lethal" Preterminal Protein Mutant Adenoviruses

Jerome Schaack,1,2,3,4,* William Y. Ho,5 Shawna Tolman,1 Elizabeth Ullyat,1 Xiaoling Guo,1 Nina Frank,1 Paul I. Freimuth,6 Dick J. Roovers,7 and John S. Sussenbach8

Department of Microbiology,1 Molecular Biology Program,2 University of Colorado Cancer Center,3 and Biomedical Sciences Program,4 University of Colorado Health Sciences Center, Denver, Colorado; Division of Clinical Research, Fred Hutchinson Cancer Research Center, Seattle, Washington5; Biology Department, Brookhaven National Laboratory, Upton, New York6; and Department of Haematology, University Hospital Utrecht,7 and Laboratory for Physiological Chemistry, Utrecht University,8 Utrecht, The Netherlands

Received 12 April 1999/Accepted 16 July 1999

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°C in HeLa-pTP cells and at 32°C and 39.5°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.

* Corresponding author. Mailing address: University of Colorado Health Sciences Center, Box B-175, 4200 E. 9th Ave., Denver, CO 80262. Phone: (303) 315-6883. Fax: (303) 315-6785. E-mail: jerry.schaack{at}uchsc.edu.

Journal of Virology, November 1999, p. 9599-9603, Vol. 73, No. 11
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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