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. Author manuscript; available in PMC: 2019 Oct 1.
Published in final edited form as: Virology. 2018 Aug 14;523:110–120. doi: 10.1016/j.virol.2018.08.003

Fig. 1. Construction of VZV gM mutants.

Fig. 1

(A) VZV gM has a 76 amino acid C-terminal domain (TMD) that contains two YXXΦ motifs, YGAV and YGYV (aa 373–376 and aa 407–410, respectively), and a dileucine (LL) motif within an acidic cluster (REDELL, aa 421 to 426) identified by linear motif scan. In all, six mutated pETVAVR fragments were generated using the recombinant vaccine Oka (rVOka) background. rVOka gM Y373A has a Y to A substitution in the YGAV sequence. rVOka gM LL425HV has a histidine-valine substituted for the native dileucines. rVOka gM Y373A+LL425HV has both of these changes. rVOka double YXXΦ has the Y373A change and a Y t o S substitution in the second YXXΦ motif. rVOka gM triple has all three changes: the Y373A, Y407S and LL425HV. The rVOka gM C-term stop mutant has two in-frame stop codons and one frame-shifted stop from aa 376. (B) The VZV genome is ~125 KB, and contains a unique long region (UL) flanked by repeats regions (long terminal repeat, TRL, and long internal repeat IRL) and a unique short region flanked by repeat regions (short internal repeat, TRS, and short terminal repeat, TRS). The VZV five cosmid system consists of five overlapping fragments of vaccine Oka strain cloned into SuperCos-1 cosmid vectors that recombine when cotransfected to form a single infectious clone of recombinant vaccine Oka strain (rVOka). ORF50 encodes VZV gM on the reverse strand of the 28506 basepair pvAvr10 cosmid at nucleotides 20198–18892. The start codons for ORF50 overlaps with the start codon for ORF51. ORF51 encodes the essential replication origin binding protein.