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Table 2 PCR primers used in this study

From: Role of sgcR3 in positive regulation of enediyne antibiotic C-1027 production of Streptomyces globisporus C-1027

Name Sequence Purpose
WAB1
WAB2
5'-GAGAATTC ACCACATGGAACATCTGCTG-3'
5'-GAAGATCT GGGAGCACGCAACTGAG-3'
Forward and reverse primers for amplifying 1.4 kbp upstream arm used in knockout of sgcR3
WCD1
WCD2
5'-AAGATCT CTGATCGGCAAGTACGGAGAC-3'
5'-AAAGCTT GGTGACAGTTCCGAGGAGGTT-3'
Forward and reverse primers for amplifying 1.4 kbp downstream arm used in knockout of sgcR3
H1
H2
5'-CAGAATTC TCTCCTACGCCCTGCTCATC-3'
5'-ACTCTAGA AGAACCGCTCGGCCTGTC-3'
Forward and reverse primers for amplifying 2,539 bp sgcR3 including its native promoter
sgcR3_FOR
sgcR3_REV
5'-GCATATG GCGGCACACGACGTGAG-3'
5'-AGGATCC CAGCTCCCCTCCTGGGAG-3'
Forward and reverse primers for amplifying 1,188 bp sgcR3 coding region
P17
P18
5'-ATGGATCC GCCCCTCGCTCATTGTCG-3'
5'-ATGAATTC CGGATCGGATGTGCTGGTCT-3'
Forward and reverse primers for amplifying 2,461 bp sgcR1R2 including its native promoter
hrdBRT1
hrdBRT2
5'-TGGTCGAGGTCATCAACAAG-3'
5'-TGGACCTCGATGACCTTCTC-3'
Forward and reverse primers used for the detection of hrdB transcripts
R1RT1
R1RT2
5'-GAAAAGTGACTCTGCCCAACGC-3'
5'-CGCTGCACATGGGAACGATC-3'
Forward and reverse primers used for the detection of sgcR1 transcripts
R2RT1
R2RT2
5'-ACCACGAACACCATCGAGGAC-3'
5'-CGGAAGATGCGGGTGAAATG-3'
Forward and reverse primers used for the detection of sgcR2 transcripts
R3RT1
R3RT2
5'-GTGGGCGAACGGGAGACGGT-3'
5'-TGCCGAGGGCCGATCAGAGG-3'
Forward and reverse primers used for the detection of sgcR3 transcripts
A1RT1
A1RT2
5'-CAGAACATATTCAAGGACTGGC-3'
5'-CACTTGTACTGTCGGGTGGTT-3'
Forward and reverse primers used for the detection of sgcA1 transcripts
C4RT1
C4RT2
5'-CTGTGGCTACGGACGAGATTG-3'
5'-CCGAACAGAACATCCCCATCT-3'
Forward and reverse primers used for the detection of sgcC4 transcripts
sgcA1p_FOR_F
sgcA1p_REV
5'-CAGCTGGAGTACGCGATGG-3'
5'-CGACAATGGCGACCTC TTTG-3'
Forward and reverse primers for amplifying 438 bp upstream of sgcA1
sgcB1p_FOR_F
sgcB1p_REV
5'-GAGGGGTTTTGAGGGGTGAA-3'
5'-GATAGGGGTCGGGGGACATC-3'
Forward and reverse primers for amplifying 496 bp upstream of sgcB1
sgcC1p_FOR_F
sgcC1p_REV
5'-GGTGCAACACCGCAGATCC-3'
5'-GAGCTGCGCGGAAGGAAG-3'
Forward and reverse primers for amplifying 481 bp upstream of sgcC1
sgcD2p_FOR_F
sgcD2p_REV
5'-CCTGGAACCATCCGCGAAAC-3'
5'-CGGTCGATCACCAGCACTTC-3'
Forward and reverse primers for amplifying 448 bp upstream of sgcD2
sgcKp_FOR_F
sgcKp_REV
5'-AGGATCATCGTCCCGTCTC-3'
5'-GAGCCAGTCGAACGACTCCA-3'
Forward and reverse primers for amplifying 471 bp upstream of sgcK
R1R2p_FOR_F
R1R2p_REV
5'-ACGATCGTGTCCGCGTAGG-3'
5'-AACGATCGATTCCGTTCAGG-3'
Forward and reverse primers for amplifying 455 bp upstream of sgcR1R2
R3p_FOR_F
R3p_REV
5'-GCCCACGCCCTCTGATCG-3'
5'-CGGGACCATGCTCCGTGTA-3'
Forward and reverse primers for amplifying 467 bp upstream of sgcR3
cagApEMSA1
cagApEMSA2
5'-CACGCTCTGTCCGTCACTCA-3'
5'-ATCCGCTCCCGAAGGTGG-3'
Forward and reverse primers for amplifying 429 bp upstream of cagA
  1. Restriction endonuclease recognition sequences introduced by these oligonucleotides are bold.