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. 2006 Nov 10;73(1):218–225. doi: 10.1128/AEM.01100-06

TABLE 1.

Primers used in this study

Category Primer Sequencea Cut site(s)
PBin-mtx1 fusion BinmutF GGAGATGAAGAAACCATGGGCGATTCAAAAGAC NcoI
BinmutR GTCTTTTGAATCGCCCATGGTTTCTTCATCTCC NcoI
mtx1F CCATGGCTTCACCTAATTCTCCAAAAG NcoI
mtx1R CCATGGTCATTACTATCTAGGTTCTACAC NcoI
mtx1 mutants MtxF CCGGAATTCCCGGATCCTCACCTAATTCTCCAAAAGAT EcoRI, BamHI
MtxR GGCCCGGGATCCTATCTAGGTTCTACACCTAATG SmaI, BamHI
MutLeuF GGATTCTAAAGGTTTGATACTAGATTTAG
MutGlyF GGATTCTAAAGGTGGTATACTAGATTTAG
MutArgF GGATTCTAAAGGTCGTATACTAGATTTAG
MutKKF GGAAATAATATGGATAAGAAAGGTAAGATACTAGATTTAG
MutLeuR CTAAATCTAGTATCAAACCTTTAGAATCC
MutGlyR CTAAATCTAGTATACCACCTTTAGAATCC
MutArgR CTAAATCTAGTATACGACCTTTAGAATCC
MutKKR CTAAATCTAGTATCTTACCTTTCTTATCCATATTATTTCC
Mtx2 production mtx2N15TF GAAGGATCCATGGAATTGTTTGTAAACGGAAGTATATATACG BamHI, NcoI
mtx2N15TR GGGCTCGAGTTATTTAAAAGAAATTTCTTTAACATCTATTA XhoI
Domain II variants A GGATCCTTTTGATGGTAGTTTTCGTGGA BamHI
B GACGTCAGTATAAATGGTTATACTATTAAG AatII
C AATTATTGGTCAGGGCATCAAA
D ATATAAAGGTGAAGATAATGTTCT
E CTGTTTGTCCTTGATGGAACG
F CAGCATTGTAACATGACTCAATC
G TTGAGAGCTCCAACGTTTTCTTGGCAGCATCGCAGTGCGGCCG SacI, NotI
H CGGCCGCACTGCGATGCTGCCAAGAAAACGTTGGAGCTCTCAA NotI, SacI
1.1 CTTAATAGTATAACCATTTATACTGACGTC(NNS)4AATTTATTGGTCAGGGCATCAAA AatII
1.2 CTTAATAGTATAACCATTTATACTGACGTC(NNS)3AATTTATTGGTCAGGGCATCAAA AatII
1.3 CTTAATAGTATAACCATTTATACTGACGTC(NNS)5AATTTATTGGTCAGGGCATCAAA AatII
2.1 AGAACATTATCTTCACCTTTATAT(NNS)13CTGTTTGTCCTTGATGGAACG
2.2 AGAACATTATCTTCACCTTTATAT(NNS)12CTGTTTGTCCTTGATGGAACG
2.3 AGAACATTATCTTCACCTTTATAT(NNS)14CTGTTTGTCCTTGATGGAACG
3.1 GATTGAGTCATGTTACAATGCTG(NNS)9TTGAGAGCTCCAACGTTTTCT
3.2 GATTGAGTCATGTTACAATGCTG(NNS)8TTGAGAGCTCCAACGTTTTCT
3.3 GATTGAGTCATGTTACAATGCTG(NNS)10TTGAGAGCTCCAACGTTTTCT
cry1Aa construction N-termF GGATCCTAAGTAGATTGTTAACACCCTG BamHI
C-term F GACGTCCATAGAGGCTTTAATTATTGGTC AatII
C-termR GGATCCCTATTCCTCCATAAGGAGTAATTC BamHI
a

The sequences of the restriction enzymes noted in the fourth column are underlined.