Oligonucleotides for PCR and site-drected mutagenesis

Oligonucleotides to generate HelB-PhoA/LacZ fusion proteins

HelB P1 5' CGGTCGACGCCAGGATCTCGCCCTGCGGGC 3'

HelB C2 5' TCGTCGACGCCAGCGCCTTGATCTGCACCA 3'

HelB P2 5' AGGTCGACAGCCAGAGATAGGCGGGCGCGG 3'

HelB C3 5' ACGTCGACCCCCCCCGCTTCAGCCCCACCG 3'

HelB P3 5' CCGTCGACACTTCGATCGCCAGGCCCTCGG 3'

Oligonucleotides to generate HelC-PhoA/LacZ fusion proteins

HelC P1 5' CCGTCGACACGTGGACATAGATCACCTTGA 3'

HelC C2 5' CCGTCGACGCCAGCGCCGAGACGTGATGAC 3'

HelC P2 5' ACGTCGACTCCCATTCCCACCAGGTTCCCC 3'

HelC C3 5' TCGTCGACGCGGCGGTATCGGGGTTCTCGA 3'

HelC P3A 5' AGCGTCGTCGACTGATGCAGGCCCTCAGAAG 3'

HelC P3B 5' GCGTCGACACATCCGCAATATGCTCTTCCT 3'

Oligonucleotides to generate Ccl1-PhoA/LacZ fusion proteins

Ccl1 P1A 5' GGCCGTCGACCCCGACCAGCCCTTTTGCGC 3'

Ccl1 P1B 5' CATCGTCGACTCATGGTTGCCCCAGACCCC 3'

Ccl1 C1 5' CACCGTCGACACCCGCGCCCGCAGCCGCTC 3'

Ccl1 P2 5' GACAGTCGACACGTACAGGAACGGCGGATG 3'

Ccl1 C2 5' CAAGGTCGACGGCCGGACCCAGCGCGCCCA 3'

Ccl1 P3B 5' ACCCCGGTCGACGGGTCATTGGCGAAACTGTG 3'

Ccl1 C3 5' CACGTTCTCGAGTCACGTCGACGATTCGCGGCTGACCATGGA 3'

Ccl P4 5' GGTAGTCGACTTTTCGAAGAACGGCGCCCC 3'

Ccl1 C4 5' GGGCGTCGACGGCCGCACCAGCTTGCCGAG 3'

Ccl1 P5 5' CGCCGTCGACGGCCGCCCCGTCGCCATCAC 3'

Ccl1 C5 5' GCCCGTCGACCCATGCGCGAAGGCCTTGCC 3'

Ccl1 P6 5' CGCCGTCGACCAGTTCGCGAAAGGCTTGAC 3'

Ccl1 C6 5' GAGCGTCGACCACTCCGCCGCAACAGCATC 3'

Oligonucleotides to generate CcsA-PhoA fusion proteins

CCSA N-terminus 5'GGACGTCATATGAACACTCTGCACGTCAACATC 3'

CCSA C1A 5' GGCGACCGACCCGGGCACCACCAGATCTCGGTTG 3'

CCSA C1B 5' GGCCAACCCAGCCTGCCCGACAAGATCTCCGAAC 3'

CCSA P2A 5' AACCCACAGCGGGCGGTACTGTAGATCTCGCAAC 3'

CCSA P2B 5' GCCGAGGCTGACCACCGACACGAGATCTGGCAGC 3'

CCSA C2 5' GATCGTCGTTCGATAGGCGATCAGATCTAGGGTC 3'

CCSA P3 5' CCACGCAACAAAAGAAACCGTCAGATCTGGATCC 3'

CCSA C3 5' GTCCCGCCATCCCGCGGTCGACAGATCTTGTAGA 3'

CCSA C-terminus 5' AACCTCGAGATCTTCCACTCCCGCATACGAATGCAGACC 3'

Oligonucleotides to generate site-directed mutants

HelC H58G 5' CAGCGTCGCCGCCGGCACNNHGACATAGATCACCTT 3'

HelC H58M 5' GTGATCTATGTCATGGTTCCGGCGGCG 3'

HelC H58S/T/V 5' CAGCGTCGCCGCCGGCACNNHGACATAGATCACCTT 3'

Ccl1 D242E/P243A 5' CGAGGCGTTTTCCACCGMSTCSCAGAACCAGAAGCCGCC 3'

HELC W117L/G118A/D124E 5' CGAGGTCAACCGCGGSTCCCATTCCCACCAGGTTSCCMACATCG 3'

Nucleotide code

S=G or C
M=A or C
H=A, C, or T
N=A, C, G, or T

Amino Acid Residue code

A=Alanine
D=Aspartate
E=Glutamate
G=Glycine
H=Histidine
L=Leucine
M=Methionine
P=Proline
S=Serine
T=Threonine
V=Valine
W=Tryptophan

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Topology of HelBCD, Ccl1, and CcsA proteins

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