TABLE 1.
Primers used in this study
F, forward; R, reverse.
| Constructa | Primers | |
|---|---|---|
| CerS2 | F | 5′- CCGAATTCCTCCAGACCTTGTATGATTACTTCTGb |
| R | 5′- CCCTCGAGTCAGTCATTCTTACGATGGTTGTTATTGAGGATGGGGb | |
| CerS5 | F | 5′- CCGAATTCGCGACAGCAGCGCAGGGACCCb |
| R | 5′- CCCTCGAGTTACTCTTCAGCCCAGTAGCTGCCTCCCATb | |
| C51–221:C2 | F | 5′- TGTTTGTGCATCACGTGGCCACCATCATTCTCb |
| R | 5′- TGGTGGCCACGTGATGCACAAACATGATCAGGAAGb | |
| C51–214:C2 | F | 5′- CAGTTTACAGACATTAAAAGAAAGGACTTCAAGGAACAGATCATCCACb |
| R | 5′- GTTCCTTGAAGTCCTTTCTTTTAATGTCTGTAAACb | |
| C51–191:C2 | F | 5′- TTCAAGTGGGCTTTATCACTATTATATGATTGAACTTTCCTTCTACTGGTCCb |
| R | 5′- AAGTTCAATCATATAATAGTGATAAAGCCCACTTb | |
| C2:C5131–392 | F | 5′- TCGCCGCCGCAATCAGGACAAGCCCCCAACGCb |
| R | 5′- GTTGGGGGCTTGTCCTGATTGCGGCGGCGACGGb | |
| C2:C5152–392 | F | 5′- AGCCAGCTGGAGATTCACATTTTACTTATGTATATTCTGCTATGGAATTAGAb |
| R | 5′- ATATACATAAGTAAAATGTGAATCTCCAGCTGGCTTCb | |
| C2:C5159–392 | F | 5′- GCCTTCATTGCCGGCATTAGATTTCTCTGGTCGTCACCTTb |
| R | 5′- CCAGAGAAATCTAATGCCGGCAATGAAGGCb | |
| C2:C5163–392 | F | 5′- TCGTCACCTTGGTTCTGGGAb |
| R | 5′- CTGTCGGATGTCCCAGAACCAAGGTGACGACCAAATGACGGCCATGCCGGCAAb | |
| C2:C5166–392 | F | 5′- CATTGTGGATAAACCTTGGTTCTGGGACATCCGACAb |
| R | 5′- ATGTCCCAGAACCAAGGTTTATCCACAATGACGGCCATGb | |
| C51–340:C2 | F | 5′- TTTGAAAGCCTTGATCGGAAAGCTGGTAGAAGATGAACGb |
| R | 5′- TCTACCAGCTTTCCGATCAAGGCTTTCAAAGCAATCCGb | |
| C51–331:C2 | F | 5′- GCGCATGGCCCACAAGb |
| R | 5′-TTTCCAGTTATGAACTTGTGGGCCATGCGCAAAATTAGGTAGGACCAGATGACATGCAb | |
| C51–309:C2 | F | 5′- CGGGCCTTATGCTTCATATTACTTCTTCAATTCCATGATGGGAb |
| R | 5′- TTGAAGAAGTAATATGAAGCATAAGGCCCGATTATCb | |
| C51–296:C2 | F | 5′- ACGACCCTGGTGTACCCACTGGAb |
| R | 5′- ATAGAGCTCCAGTGGGTACACCAGGGTCGTGTTCAGAATCCAGAATGb | |
| C2:C5152–340:C2 | F | 5′- GATTCACATTTTACTTATGTATATTCTGCTAT |
| R | 5′- AGCAGAATATACATAAGTAAAATGTGAATCTCCAGCTGGC | |
| C2:C5152–309:C2 and C2:C5159–309:C2 | F | 5′- GATAATCGGGCCTTATGCTTCATATTACTTCTTCAATTCCATGc |
| R | 5′- GGTACCGGGCCCCCCCTCGAGTCc | |
| C5:C2144–332:C5 | F | 5′- GAAAGCATGTGGAGATTCACATTTTATCTGATTGCCTTCATTGCCGGCd |
| R | 5′-GATCATCCTTCGATACCTTTCCCCTAGTTATGAACTTGTGGGCCATGCd | |
| C5:C2151–301:C5 | F | 5′- CATTTTATTTATGTATATTCTGCTATGGAATGGCCGTCATTGTGGATAAACCd |
| R | 5′- GCAGGCCATTGAGGAGCCACCAGCCAAAGAAGGCAGGATAGAGCd |
a The terminology used for each construct is given in Fig. 2.
b External and matching internal primers were used to amplify the desired regions from CerS2 or CerS5 templates.
c Used to amplify desired regions from C51–309:C2 and subsequently inserted into C2:C5152–392 or C2:C5159–392 by restriction free cloning.
d Used to amplify regions from CerS2 and subsequently inserted into CerS5 by restriction free cloning.