Table 1 Summary of selection for "Specialist" and "Generalist" polymerases
From: A combined in vitro / in vivo selection for polymerases with novel promoter specificities
A. Selection of "Specialist" (T3 like polymerase) | |||||
|---|---|---|---|---|---|
§Number of variants | Polymerase sequence* | Promoter sequence (-11 to -8) | |||
746 | 747 | 748 | |||
Round – 1 | 4 | R (CGA) | L (TTG) | D (GAT) | CACT |
1 | R (CGT) | L (TTA) | Q (CAG) | CACT | |
1 | R (CGG) | L (TTA) | S (TCT) | CACT | |
1 | R (CGG) | C (TGT) | S (TCT) | CACT | |
1 | R (CGC) | P (CCC) | E(GAA) | CACT | |
1 | R (AGG) | T (ACA) | N(AAC) | CACT | |
Round – 2 | 17 | R (CGA) | L (TTA) | D (GAT) | CACT |
3 | R (CGA) | L (TTG) | Q (CAG) | CACT | |
Round – 3 | 16 | R (CGA) | L (TTA) | D (GAT) | CACT |
* wild type sequence: | R (CGC) | L (TTG) | N (AAC) | GACT | |
B. Selection of the "Generalist" polymerase | |||||
§Number of variants | Polymerase sequence* | Promoter sequence (-11 to -8) | |||
748 | 756 | 758 | |||
Round – 3 | |||||
R3–9 | 3 | N(AAT) | R (CGT) | C (TGC) | GACT(wt) |
R3–32 | 3 | N(AAT) | R (CGT) | C (TGC) | GACG |
R3–26 | 1 | N(AAT) | R (CGT) | C (TGC) | TGTA |
R3–17 | 1 | A (GCC) | M (ATG) | S (AGT) | GGTA |
R3–5 | 1 | F (TTC) | G (GGC) | I (ATA) | G CTA |
R3–29 | 1 | T (ACG) | K(AAG) | Q (CAA) | GACT |
Round – 4 | |||||
R4–14 | 7 | N(AAT) | R (CGT) | C (TGC) | GACT |
R4–16 | 3 | N(AAT) | R (CGT) | C (TGC) | GTTA |
R4–17 | 2 | N(AAT) | R (CGT) | C (TGC) | GTCA |
ppC6 | 2 | N(AAT) | R (CGT) | C (TGC) | GATA |
R4–11 | 3 | N(AAT) | R (CGA) | S (AGT) | GACT |
* wild type sequence | N (AAC) | R (CGC) | Q (CAG) | GACT | |