Data from:
Randell JC, Komazin G, Jiang C, Hwang CB, Coen DM. 2005. "Effects of substitutions of arginine residues on the basic surface of herpes simplex virus UL42 support a role for DNA binding in processive DNA synthesis." J. Virol 79(18):12025-34. (Pubmed)
Supplementary Table 1.
Primers and Restriction Endonucleases for Site-Directed Mutagenesis of UL42
| Amino acid change(s) | Nucleotide change(s) | Primer pairs (Round 1)a | Primer pair (Round 2) | Restriction sites for cloning |
|---|---|---|---|---|
| R113A | C337G/G338C | 5’AGCGTGTTTGCCGCCAACCAG 5’GGGGAACCAGCACCAC 5’CTGGTTGGCGGCAAACACGCT 5’CGGGATGGAGGGAAGG |
5’GGGGAACCAGCACCA 5’CGGGATGGAGGGAAGGC |
BamH I, Nhe I |
| R182A | A544G/G545C | 5’CGCCTGACGGCGCCGCAGCTC 5’GGGTTTTCCCAGTCAC 5’GAGCTGCGGCGCCGTCAGGCG 5’CGGGATGGAGGGAAGG |
5’GGGTTTTCCCAGTCAC 5’CGGGATGGAGGGAAGG |
Nhe I, Sph I |
| R279A | C835G/G836C | 5’GCGGTGCTCGCGCGACTGCAG 5’GGGTTTTCCCAGTCAC 5’CTGCAGTCGCGCGAGCACCGC 5’CCCCGTTTCGCACGCTGG |
5’GGGTTTTCCCAGTCAC 5’CCCCGTTTCGCACGCTGG |
Sph I, Hind III |
| R280A | C838G/G839C | 5’GTGCTCCGGGCACTGCAGGTC 5’GGGTTTTCCCAGTCAC 5’GACCTGCAGTGCCCGGAGCAC 5’CCCCGTTTCGCACGCTGG |
5’GGGTTTTCCCAGTCAC 5’CCCCGTTTCGCACGCTGG |
Sph I, Hind III |
| R279A/R280A | C835G/G836C/ C838G/G839C |
5’GCGGTGCTCGCGGCACTGCAGGTCGGCGGG 5’GGGTTTTCCCAGTCAC 5’CCCGCCGACCTGCAGTGCCGCGAGCACCGC 5’CCCCGTTTCGCACGCTGG |
5’GGGTTTTCCCAGTCAC 5’CCCCGTTTCGCACGCTGG |
Sph I, Hind II |
a. Altered codons shown in bold