Table 3 Primers used for PCR amplification and cloning of recombinant forms of AaET, AaSPVI, AaSPVII, and AaLT, without (Zymogen), or with, an enterokinase cleavage site (Mek)
Protease | Primer | Primer Sequence |
|---|---|---|
AaET-Zymogen | AR1-ET-ORF-F | 5'-AAAAAAACATATG AACCAATTTCTCTTTGTCAG-3' |
AR2-ET-ORF-R | 5'-AAAAAAAAAGCTT ATTAAACCTCGGAAACCTCTCG-3' | |
AaSPVI-Zymogen | AR17-5G1-3714-F | 5'-AAAAAAACATATG GTTCGCATCATTCTTATTCT-3' |
AR18-5G1-3714-R | 5'-AAAAAAAAAGCTT ATTACAATCCACTGACCTCCTGCACCC-3' | |
AaSPVII-Zymogen | AR15-CxLTA1-ORF-F | 5'-AAAAAAACATATG GCTCGTATCATCCTTCTGTT-3' |
AR16-CxLTA1-ORF-R | 5'-AAAAAAAAAGCTT ATTAAACTCCACTGACTTCGGCC-3' | |
AaLT-Zymogen | AR4-LT-ORF-F | 5'-AAAAAAGCATATG TTCACTTCAACGGTGGT-3' |
AR5-LT-ORF-R | 5'-AAAAAAGCTCGAG TTATTACAGTCCAGTCTTCTGCTTGAT-3' | |
AaET-Mek | AR28-ET-M-pET-EK-F | 5'-AAAAACATATG AACAACAACCTCGGCGATGACGATGACAAG ATCGTTGGCGGATTCCAGAT-3' |
AR2-ET-ORF-R | 5'-AAAAAAAAAGCTT ATTAAACCTCGGAAACCTCTCG-3' | |
AaSPVI-Mek | AR27-5G1-M-pET-EK-F | 5'-AAAAACATATG AACAACAACCTCGGCGATGACGATGACAAG ATTGTTGGTGGCTTTGAAGT-3' |
AR18-5G1-3714-R | 5'-AAAAAAAAAGCTT ATTACAATCCACTGACCTCCTGCACCC-3' | |
AaSPVII-Mek | AR29-CxLT-M-pET-EK-F | 5'-AAAAACATATG AACAACAACCTCGGCGATGACGATGACAAG GTCGTCGGCGGTTATGAAGT-3' |
AR16-CxLTA1-ORF-R | 5'-AAAAAAAAAGCTT ATTAAACTCCACTGACTTCGGCC-3' | |
AaLT-Mek | AR30-LT-M-pET-EK-F | 5'-AAAAACATATG AACAACAACCTCGGCGATGACGATGACAAG GTAGTAAACGGACAAACGGC-3' |
AR5-LT-ORF-R | 5'-AAAAAAGCTCGAG TTATTACAGTCCAGTCTTCTGCTTGAT-3' |