Figure 1.

Construction and analysis of TNsubUL69. (A) A substitution mutant was produced in which a 2,144-bp segment within the UL69 coding region extending from sequence position 98290 to 100433 (AD169 sequence numbers) was replaced with the EGFP coding region. (B) Human fibroblasts were infected with wild-type virus (TNwt), mutant virus grown in the presence of helper retrovirus (TNsubUL69^+pUL69), or mutant virus grown for one cycle in the absence of helper retrovirus (TNsubUL69^−pUL69), and UL69 protein was assayed by Western blot using a UL69-specific monoclonal antibody (α-UL69) at 72 h after infection. The late protein, pp28, was assayed as a control by using a pp28-specific monoclonal antibody (α-pp28). The expression of late viral gene products is reduced in mutant virus-infected cells. (C) Virus particles were partially purified and virion proteins were assayed by Western blot.