Data From:
Loregian A, Appleton BA, Hogle JM, Coen DM.2004. "Residues of human cytomegalovirus DNA polymerase catalytic subunit UL54 that are necessary and sufficient for interaction with the accessory protein UL44." J. Virol. 78:158-67. (Pubmed)
Supplementary material
List of primer pairs used for creation of UL44 constructs.
Construct | PCR round | Forward primer | Reverse primer |
---|---|---|---|
GST-UL44 full-length | First round | 5’-CTGGAAGTTCTGTTCCAGG GGCCCCTCGAGATGGATCGCAAGACGCG-3’ |
5’-AGCTGGGTCACGCGTCTAGCC GCACTTTTGCTTCTT-3’ |
Second round | 5’-GGGGACAAGTTTGTACAA AAAAGCAGGCTTACTGGAAGTTCTGTTCCAG-3’ |
5’-GGGGACCACTTTGTACAAGAA AGCTGGGTCACGCGTCTA-3’ |
|
GST-UL44DC290 | First round | 5’-CTGGAAGTTCTGTTCCAGG GGCCCCTCGAGATGGATCGCAAGACGCG-3’ |
5’-AGCTGGGTCACGCGTCTAACG CGACTTGCCGCTGTT-3’ |
Second round | 5’-GGGGACAAGTTTGTACAA AAAAGCAGGCTTACTGGAAGTTCTGTTCCAG-3’ |
5’-GGGGACCACTTTGTACAAGAA AGCTGGGTCACGCGTCTA-3’ |
List of primer pairs used for creation of UL54 mutants.
Mutant | Forward primer | Reverse primer |
---|---|---|
DCC | 5’–AGTGTCAAGGCGCACGAATG AGAAACAGC–3’ |
5’–GCTGTTTCTCATTCGTGCGCC TTGACACT–3’ |
C1241A | 5’-gtcaaggcgcacgaagcctgt tgagaaacagcg-3’ |
5’-cgctgtttctcaacaggcttc gtgcgccttgac–3’ |
C1242A | 5’-ggcgcacgaatgcgcttgag aaacagcggcg-3’ |
5’–cgccgctgtttctcaagcgca ttcgtgcgcc–3’ |
C1241A-C1242A | 5’-gtgtcaaggcgcacgaagccg cttgagaaacagcggcg-3’ |
5’–cgccgctgtttctcaagcggc ttcgtgcgccttgacac–3’ |
E1240A | 5’-gtgtcaaggcgcacgcatgct gttgagaaacag-3’ |
5’–ctgtttctcaacagcatgcgt gcgccttgacac–3’ |
H1239A | 5’-ccgtacagtgtcaaggcggcc gaatgctgttgagaaac-3’ |
5’–gtttctcaacagcattcggcc gccttgacactgtacgg–3’ |
K1237A | 5’-ctgccgtacagtgtcgcggcg cacgaatgctg-3’ |
5’–cagcattcgtgcgccgcgac actgtacggcag–3’ |
V1236A |
5’-ctgccgtacagtgccaaggcg cacgaatgc-3’ |
5’–gcattcgtgcgccttggcact gtacggcag–3’ |
S1235A | 5’-gcttttctgccgtacgctgtc aaggcgcacgaatgc-3’ |
5’–gcattcgtgcgccttgacagc gtacggcagaaaagc–3’ |
Y1234A | 5’-ccggcttttctgccggccagt gtcaaggcgcac-3’ |
5’–gtgcgccttgacactggccg gcagaaaagccgg–3’ |
P1233A | 5’-gagccggcttttctggcgtac agtgtcaaggcg-3’ |
5’–cgccttgacactgtacgcca gaaaagccggctc–3’ |
L1232A | 5’-cttggagccggcttttgcgcc gtacagtgtcaaggc-3’ |
5’–gccttgacactgtacggcgc aaaagccggctccaag–3’ |
F1231A | 5’-cacttggagccggctgctctg ccgtacagtgtc-3’ |
5’–gacactgtacggcagagcag ccggctccaagtg–3’ |
F1231Y | 5’–CACTTGGAGCCGGCTTATCTG CCGTACAGTGTC–3’ |
5’–GACACTGTACGGCAGATAAG CCGGCTCCAAGTG–3’ |
P1229A | 5’-cgcttgcacttggaggcggct tttctgccgtac-3’ |
5’–gtacggcagaaaagccgcct ccaagtgcaagcg–3’ |
E1228A | 5’-gcgcttgcacttggcgccggc ttttctgc-3’ |
5’–gcagaaaagccggcgccaag tgcaagcgc–3’ |
L1227A | 5’-cggcgcttgcacgcggagccg gcttttctg-3’ |
5’–cagaaaagccggctccgcgt gcaagcgccg–3’ |
H1226A | 5’-ctgccgcggcgcttggccttg gagccggc-3’ |
5’–gccggctccaaggccaagcg ccgcggcag–3’ |
L1225A | 5’-gctgccgcggcgcgcgcactt ggagccg-3’ |
5’–cggctccaagtgcgcgcgcc gcggcagc–3’ |
R1224A | 5’-gtgctgccgcgggccttgcac ttggagccg-3’ |
5’–cggctccaagtgcaaggccc gcggcagcac–3’ |
R1223A | 5’-cacatggtgctgccggcgcgc ttgcacttggag-3’ |
5’–ctccaagtgcaagcgcgccg gcagcaccatgtg–3’ |
P1222A | 5’-cacatggtgctggcgcggcgc ttgcacttg-3’ |
5’–caagtgcaagcgccgcgcca gcaccatgtg–3’ |
L1221A | 5’-gtttttgcacatggtggcgcc gcggcgcttgc-3’ |
5’–gcaagcgccgcggcgccacc atgtgcaaaaac–3’ |
F1217A | 5’-gcgcgcaaggacaaggctttg cacatggtgctgc-3’ |
5’-gcagcaccatgtgcaaagcc ttgtccttgcgcgc-3’ |