FIG. 1.
Construction of recombinant VZV gB-36. (A) The prototype VZV genome (Dumas strain) is 124,884 bp in length (top line), and the position of ORF31, which encodes gB, is shown. The four overlapping cosmids used to generate infectious virus are depicted (second and third lines), and the nucleotide coordinates for the MstII B cosmid, which contains ORF31, are given. ORF31 (nucleotides 57,008 to 59,611) is expanded in the fourth line, and the direction of its translation is indicated (arrow). The shaded area represents the region of ORF31 predicted to encode the cytoplasmic domain of gB. The codon CGC at amino acid position 833 (beginning at nucleotide 59,504) within cosmid MstII B was mutated to TGA, changing it from an arginine (R) to a stop codon. (B) C-terminal amino acid sequences of native VZV gB and VZV gB-36. The numbers on top are the coordinates within gB, and the asterisk indicates the position of the stop codon that was introduced to generate the truncated form of gB. The underlined sequences were previously shown to be required for the ER-to-Golgi transport of gB, and the bold sequences are consensus YXXφ motifs.