Table 1. GPCMV Glycoprotein Genes, predicted size, homology and knockout site.
| GPCMV Gene (co-ordinates) | Glycoprotein (predicted size) | Signal peptide predicted | Glycosylation Sites | % Identity with HCMV (BLAST) | Site of ORF Knockout |
|---|---|---|---|---|---|
| GP55 (c94164-96869) | gB (901 aa/ 102.2 kDa) | yes | 32 (O-linked) 15 (N-linked) | 45% | codon 528 insertion |
| GP73 (117644–118045) | gN (134 aa/ 14 kDa) | yes | 17 (O-linked) 2 (N-linked) | 44% | codon 71 insertion |
| GP74 (c117992-119105) | gO (370 aa / 41.8 kDa) | 13(O-linked) 13(N-linked) | 27% | codon 110 insertion | |
| GP75 (c119553-121724) | gH (724 aa/ 81.8 kDa) | yes | 10 (O-linked) 9 (N-linked) | 29% | codon 200 insertion/del |
| GP100 (c157482-158531) | gM (349 aa/ 39.7 kDa) | yes | 0 (O-linked) 2 (N-linked) | 52% | codon 170 insertion |
| GP115 (c180216-180992) | gL (258 aa/ 29.7 kDa) | yes | 3 (O-linked) 3 (N-linked) | 42% | codon 45 insertion |
c = complement DNA strand coding, aa = amino acids.
Co-ordinates based on complete GPCMV (22122 strain) sequence (GenBank: AB592928.1); Kanai et al. [10]. Percentage identity determined by BLAST analysis of GPCMV against HCMV Towne strain. Predicted protein size is based on complete protein predicted sequence and calculated using MacVector.
Signal peptide sequence predicted by web based programs. See S4 Fig
Post translational glycosylation predicted based on web programs: NetOGlyc 4.0 Server (http://www.cbs.dtu.dk/services/NetOGlyc/) for O-glycosylation; and NetNGlyc 1.0 Server (http://www.cbs.dtu.dk/services/NetNGlyc/) for N- glycosylation. Total predicted number of N-glycosylation or O-glycosylation sites per glycoprotein are indicated.
Insertion site of the kanamycin (Km) cassette to knockout the various coding sequences was carried out using convenient restriction sites (see materials and methods). The insertion of the Km cassette disrupts the ORF at the specified codon.