TABLE 1.
Expression plasmids generated for this study
| Plasmid | Protein expressed | Insert in the expression plasmid pCAGGS |
|---|---|---|
| pAZB2 | gB2 | Nucleotides 56219 to 54368 and 54617 to 53374 from HSV-2(333)a were amplified by PCR, joined by the unique SphI site in the overlapping regions, and cloned into the EcoRI and KpnI restriction sites |
| pAZD2 | gD2 | PCR amplification of nucleotides 140992 to 142323 from HSV-2(333)a cloned into the EcoRI and KpnI restriction sites |
| pAZH2 | gH2 | Nucleotides 46608 to 45131 and 45157 to 43933 from HSV-2(333)a were amplified by PCR, joined by the unique XhoI site in the overlapping regions, and cloned into the EcoRI and KpnI restriction sites |
| pAZL2 | gL2 | PCR amplification of nucleotides 9382 to 10225 from HSV-2(333)a cloned into EcoRI and KpnI restriction sites |
| pAZCH5 | gD2(1-66)/gD1(67-369)b | Nucleotides 140992 to 141288 from HSV-2(333)a and nucleotides 138692 to 139626 from HSV-1(KOS)c were amplified by PCR and combined in a final PCR by using only the outside primers |
| pAZCH1 | gD2(1-98)/gD1(99-369) | gD2(1-98) was joined to gD1(99-369) by a natural restriction site (Van91I) |
| pAZCH16 | gD1(1-66)/gD2(67-368) | Nucleotides 138398 to 138691 from HSV-1(KOS)c and nucleotides 141289 to 142323 from HSV-2(333)a were amplified by PCR and combined in a final PCR using only the outside primers |
| pAZCH35 | gD1(1-98)/gD2(99-368) | gD1(1-98) was joined to gD2(99-368) by a natural restriction site (Van91I) |
| pAZMUT1.1 | gD1(A7P) | Codon GCC was changed to CCC, causing an Ala→Pro substitution in position 7 |
| pAZMUT2.1 | gD1(A42P,G43S) | Codons GCG and GGC were changed to CCG and AGC, causing Ala→Pro and Gly→Ser substitutions in positions 42 and 43 |
| pAZMUT3.1 | gD1(P45E) | Codon CCG was changed to GAG, causing a Pro→Glu substitution in position 45 |
| pAZMUT4.1 | gD1(A7P,D21N) | Codons GCC and GAC were changed to CCC and AAC, causing Ala→Pro and Asp→Asn substitutions in positions 7 and 21 |
| pAZMUT5.1 | gD1(A7P,P45E) | Codons GCC and CCG were changed to CCC and GAG, causing Ala→Pro and Pro→Glu substitutions in positions 7 and 45 |