Figure 1.

Schematic representation of viral and plasmid DNA sequence arrangements. Line 1, A linear representation of the HSV-1 genome. The rectangles represent the inverted repeats flanking the unique sequences (U_L and U_S represented by thin lines). Line 2, pRB5709, a 4.5-kb XhoI/NotI fragment of HSV-1(F) containing the U_L33, U_L34, U_L35, and portion of U_L32 and U_L36 genes. The arrowheads indicate the direction of the transcription; solid dotted box indicates the putative transmembrane domain of the U_L34 gene product. Line 3, DNA sequences of U_L31 coding domains showing relevant restriction endonucleases sites used for construction of deletion mutants. Line 4, From left to right, pRB5710, pRB5711, and pRB5712. pRB5710 was derived by excision of a XcmI fragment in U_L34, blunt-ending, and religation to produce a frame shift after the first two codons of U_L34. pRB5711 was made by direct religation of the plasmid pRB5709 after removal of a XcmI fragment. This caused an in-frame deletion of codons 3–119. In pRB5712 the pRB5709 plasmid was cleaved with AflII, blunt-ended, and religated. In addition, a stop codon, and a unique restriction site (PacI), was introduced immediately after codon 244 of U_L34. Lines 5–7, The transfer plasmids pRB5713, pRB5714, and pRB5715 were constructed by cloning the XhoI/NotI fragment from pRB5710, pRB5711, and pRB5712, respectively, into the transfer vector (pRB5708) at XhoI/NotI sites.