Loregian et al (2004) A

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’