Table 5. Sequences of primers used in PCR experiments The nucleotides corresponding to the coding sequences are in capital letters, the Kozak consensus sequence is shown in bold, and the added restriction sites are underlined.
| Primer | Sequence | Restriction site | Plasmid | Protein expressed |
|---|---|---|---|---|
| Preparation of SEDT3 protein expression vectors | ||||
| #1 | taaggtaccgccaccATGGGTAAGAAGAGTCGAGTG | KpnI | pEF6/Myc-His A | C-terminal His6-tagged rat SEDT3 protein |
| #2* | taagcggccgCCAGAGTCGCTCCTTCACCGC | NotI | ||
| #3 | taaggtaccgccaccATGGGTAAGAAGAGTCGAGTAA | KpnI | pEF6/Myc-His A | C-terminal His6-tagged rat SEDT3 protein |
| #4* | taagcggccgCCACTCCTTAACTCCAGCAGTG | NotI | ||
| Preparation of cofilin-1 and profilin-1 expression vectors | ||||
| #5 | tatacatATGGCCTCCGGTGTGGCTG | NdeI | pCOLD I | N-terminal His6-tagged human cofilin-1 |
| #6 | tataaagcTTATCACAAAGGCTTGCCCTCCAG | HindIII | ||
| #7 | tatacatATGGCCGGGTGGAACGCC | NdeI | pCOLD I | N-terminal His6-tagged human profilin-1 |
| #8 | tataaagcTTATCAGTACTGGGAACGCCGAAG | HindIII | ||
| Preparation of S-adenosyl-L-homocysteine nucleosidase and andenine deaminase expression vectors | ||||
| #9 | tatacatATGAAAATCGGCATCATTGGTG | NdeI | pCOLD I | N-terminal His6-tagged E. coli SAH nucleosidase |
| #10 | tataaagcTTAGCCATGTGCAAGTTTCTGC | HindIII | ||
| #11 | tataggtaccTTGAATAAAGAAGCGCTAGTCAAT | KpnI | pCOLD I | N-terminal His6-tagged B. subtilis andenine deaminase |
| #12 | tataggatccTTATTGCAGTGATATGTGTTGAAAT | BamHI |
*Original STOP codons were replaced by CCA (shown in italic and underlined) to allow C-terminal HisTag translation.