TABLE 1

Construction of chimeras: primers and templates

Sequences of primers
PrimerFunctionSequence
SEC9-+15SEC9 5′ end XhoI siteCAG TAG CTC GAG AGA ATG GGA TTA AAG AAA
SEC9-DS3SEC9 3′ at SacI siteGAT TAG GAG CTC GGT TAA GTT CA
SPO20-+17SPO20 5′ end XhoI siteCAG TAG CTC GAG ATA ATG GGG TTC AGA AAA AT
SPO20-DSSPO20 3′ end SacI siteCAG TAG GAG CTC CGA TGT TTG AAT GCA C
S9/S20-F1SEC9NT/SPO20H1 junctionCAA CTA TTT CAT CCT CCT CCT GCT GCT GGC GAG CCT CC
S20/S9-F1SPO20NT/SEC9H1 junctionCTA CTG CTT CAT CTT CTT CAG GCT GGA CAT TCT CTA AG
ANO139SEC9H1/SPO20IH junctionGAA GGG GTT GGG TGA TTT TAA AGC CAA TAT ACT ACG GTT
ANO140SPO20H1/SEC9IH junctionCGG ATT AGA AAC ATG GAC AGC AAG CAG GCT TCT ATT TTC
ANO155SEC9IH/SPO20H2 junctionTGG GCA TCT CTA AGA ATA CGC TGC CTC TGA TAT CTT TCC CGC A
ANO156SPO20IH/SEC9H2 junctionTCT TTG CCT TTT CTA GAA CAT TCT TTC TTT CCA CCC CTT GGC CAT A
ANO163Remove SEC9NTGTT CCT CTC GAG AGA ATG GGA AAA GAA GAG GAG GCT CGC C
ANO164Remove SPO20NTCTT GTT CTC GAG ATA ATG GGG CAA TAT AAC TTA GAG AAT GTC C
PCR reactions
First PCRSecond PCR
ChimeraPrimer 1Primer 2TemplatePrimer 1Primer 2Template
SSSSSEC9-+15SEC9-DS3SEC9None
PPPPSPO20-+17SPO20-DSSPO20None
SPPPSEC9-+15S9/S20-F1SEC9SEC9-+15SPO20-DSSPO20
PSSSSPO20-+17S20/S9-F1SPO20SPO20-+17SEC9-DS3SEC9
SSPPSEC9-+15ANO139SEC9SEC9-+15SPO20-DSSPO20
PPSSSPO20-+17ANO140SPO20SPO20-+17SEC9-DS3SEC9
SSSPSEC9-+15ANO155SEC9SEC9-+15SPO20-DSSPO20
PPPSSPO20-+17ANO156SPO20SPO20-+17SEC9-DS3SEC9
PSSSSEC9-+15ANO140SPPPSEC9-+15SEC9-DS3PSSS
PSPPSPO20-+17ANO139PSSSSPO20-+17SPO20-DSSPPP
SSPSSEC9-+15ANO156SSPPSEC9-+15SEC9-DS3PSSS
PPSPSPO20-+17ANO155PPSSSPO20-+17SPO20-DSSPPP
SPPSSEC9-+15ANO156SPPPSEC9-+15SEC9-DS3PSSS
PSSPSPO20-+17ANO155PSSSSPO20-+17SPO20-DSSPPP
SPSPSEC9-+15ANO155SPSSSEC9-+15SPO20-DSSPO20
PSPSSPO20-+17ANO156PSPPSPO20-+17SEC9-DS3SEC9
xSSSANO163SEC9-DS3SEC9None
xSSPANO163SPO20-DSSSSPNone
xSPSANO163SEC9-DS3SSPSNone
xSPPANO163SPO20-DSSSPPNone
xPPPANO164SPO20-DSSPO20None
xPPSANO164SEC9-DS3PPPSNone
xPSPANO164SPO20-DSPPSPNone
xPSSANO164SEC9-DS3PPSSNone