Construction and preliminary characterization of a library of "lethal" preterminal protein mutant adenoviruses

Adenoviruses containing lethal in-frame insertion mutant alleles of the pre terminal protein (pTP) gene were constructed with cell lines that express p TP. Thirty in-frame insertion mutant alleles, including 26 alleles previous ly characterized as lethal and 4 newly constructed mutant alleles, were int roduced into the viral chromosome in place of the wild-type pTP gene. The v iruses 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 newl y constructed viruses exhibited temperature sensitivity for plaque formatio n, one virus did not form plaques in the absence of complementation, seven additional mutants exhibited a greater than 10-fold reduction in plaque for mation in the absence of complementation, and another eight mutants exhibit ed stronger phenotypes than did previously characterized in-frame insertion mutants in the plaque assay. These mutant viruses offer promise for analys is of pTP functions.