Table 3.
pLexA-WRN fusion protein constructs
| Name of construct | WRN Protein coding Domaina | Primer set (Forward/reverse) | Primer sequence site (5′→3′) | pBTM116 cloning site |
|---|---|---|---|---|
| 1 | 6–220 | TH1 | BglII (GCC AGA TCT TGG AAA CAA CTG CAC) | |
| TH2 | SalI (GCC GTC GAC ACC AGC ATA AGC ATC) | BamHI-SalI | ||
| 2 | 205–412 | TH3 | EcoRI (CCG GAA TTC CTC ACT GAG GAC CAG) | |
| TH4 | SalI (GCC GTC GAC TTC TTC CTG AGA CTG) | EcoRI-SalI | ||
| 1+2 | 6–412 | TH1 | BglII (GCC AGA TCT TGG AAA CAA CTG CAC) | |
| TH4 | SalI (GCC GTC GAC TTC TTC CTG AGA CTG) | BamHI-SalI | ||
| R+ | 406–525 | TH13 | EcoRI (TGA GAA TTC GGA CAG CAG TCT CAG) | |
| TH14 | SalI (CGC GTC GAC CTT ATC ATC ATC TTC) | EcoRI-SalI | ||
| R1/2 | 406–525 | TH13 | EcoRI (TGA GAA TTC GGA CAG CAG TCT CAG) | |
| (450–476) | TH14 | SalI (CGC GTC GAC CTT ATC ATC ATC TTC) | EcoRI-SalI | |
| R− | 406–525 | TH13 | EcoRI (TGA GAA TTC GGA CAG CAG TCT CAG) | |
| (424–475) | TH14 | SalI (CGC GTC GAC CTT ATC ATC ATC TTC) | EcoRI-SalI | |
| H | 522–868 | TH15 | EcoRI (CGC GAA TTC GAT GAT GAT AAG GAC) | |
| TH16 | SalI (ATT GTC GAC CCA GAG GAC GTG ACA) | EcoRI-SalI | ||
| 3 | 861–1,062 | TH5 | EcoRI (CCG GAA TTC AGT TCT TGT CAC GTC) | |
| TH6 | SalI (GCC GTC GAC GAT GAG GCT GTG AGA) | EcoRI-SalI | ||
| 4 | 1,014–1,179 | TH7 | EcoRI (CCG GAA TTC TGG AAG GCT TTT TCC) | |
| TH8 | SalI (GCC GTC GAC TGC CAG AAT AGC TGG) | EcoRI-SalI | ||
| 5 | 1,162–1,306 | TH9 | EcoRI (CCG GAA TTC GTA GAA GCT AGG CAG) | |
| TH10 | SalI (GCC GTC GAC TGC TCG CTC CAA ATC) | EcoRI-SalI | ||
| 6 | 1,272–1,432 | TH11 | EcoRI (CCG GAA TTC CCT TTG AAG AGC ATA) | |
| TH12 | SalI (GCC GTC GAC GCT GAT TTA AGA TGC) | EcoRI-SalI | ||
| F+ | 1–1,432 | TH1 | BglII (GCC AGA TCT TGG AAA CAA CTG CAC) | |
| TH2b | SalI (GCC GTC GAC GCT GAT TTA AGA TGC) | BamHI-SalI | ||
| F1/2 | 1–1,432 | TH1 | BglII (GCC AGA TCT TGG AAA CAA CTG CAC) | |
| (450–476) | TH2b | SalI (GCC GTC GAC GCT GAT TTA AGA TGC) | BamHI-SalI | |
| F− | 1–1,432 | TH1 | BglII (GCC AGA TCT TGG AAA CAA CTG CAC) | |
| (424–475) | TH2b | SalI (GCC GTC GAC GCT GAT TTA AGA TGC) | BamHI-SalI |
a
WRNp domains are relative to amino acid positions 1–1432 of the wild-type human WRNp (accession number L76937). Internal amino acid deletions are indicated within parentheses and were constructed as described in MATERIALS AND METHODS.
b
Full-length constructs were made by insertion of the remaining 3′-WRN coding sequence into pLexA-WRN domain 1, as described in MATERIALS AND METHODS.