promoters of Streptomyces from Metabolic Engineering-2013-Siegl

来源: Streptomyces lividans TK24
功能: promoter

Summary

synthetic promoters

基本信息

定量方法 1 ml of 24-hour seed cultures of the S. lividans TK24 mutant strains was inoculated into 50 ml of TSB medium and grown in shaking flasks for 40 h at 28℃, which corresponds to the stationary phase. The culture (1 ml) was pelleted, resuspended in 900 ml of GUS buffer 1 (50 mM phosphate buffer [pH 7.0], 5 mM dithiothreitol [DTT], 0.1% Triton X-100, 1 mg/ml lysozyme) and incubated at 37℃ for 20 min. The lysates were diluted with 900 ml of GUS buffer 2 (50 mM phosphate buffer [pH 7.0], 5 mM DTT, 0.1% Triton X-100) and centrifuged at 14000 rpm at 4 ℃ for 10 min. A 500-ml sample of the supernatant was mixed with 500 ml of GUS buffer 3 (50 mM phosphate buffer [pH 7.0], 5 mM DTT, 0.1% Triton X-100 supplemented with 2 mM p-nitrophenyl-beta-D-glucuronide), and the absorption was spectrophotometrically measured at 415 nm for 30 min.
培养条件 The medium (TSB) formulations and manipulations of actinomycetes (conjugal transfer between E. coli ET12567/pUZ8002 and S. lividans TK24) were performed as previously described (Kieser et al., 2000)。Tryptone soya broth (TSB): Oxoid Tryptone Soya Broth Powder 30 g, distilled water 1000 ml.
报告基因 beta-D-glucuronidase (GUS) reporter protein
诱导剂
提交人 Lu yinhua;Zhao Wenfang

元件序列

元件名 序列 序列长度
D2
CGCGTAGGCTAAAGATGACATCTTTAACATAGGATGTGCGC
41
ermEp1
GTTGTGGGCTGGACAATCGTGCCGGTTGGTAGGATCCAGCG
43
76
AACATAGGCTATACAAAAAAACCGCAGTTTAGGATAAATAC
43
114
TCGGGGGGCTCGACACGGCCTCTAACTACTAGGATAACGCT
42
B12
CGGGATGGCTTATGAAGGATTGTCTCACTTAGGATAGAGCA
43
D1
TTTAGAGGCTCATTGATGGAATCCGAGGATAGGATAAAGTG
41
D9
CTTAGAGGCTGGTGTCAACTAGATCACCATAGGATTCACCA
41
D11
ACGCCGGGCTACAAAAATAATATCAGAACTAGGATGGACTC
41
D7
AAGAAGGGCTAAGTTGATTATGCCAGTAATAGGATAAACTA
41
82
TCCTGAGGCTAAAATTAGGATTCTTCAAGTAGGATTGTAGC
43
ermE*
GCTAGCCGCGGTCGATCTTGACGGCTGGCGAGAGGTGCGGGGAGGATCTGACCGACGCGGTCCACACGTGGCACCGCGATGCTGTTGTGGGCACAATCGTGCCGGTTGGTAGGATCCAGCG
121
C1
ACTTTCGGCTAAAAAGCAATTCATTCAATTAGGATGGAAGA
41
A7
TAAAGAGGCTTTA
15
102
TCACAGGGCTCAATGCTAAAAGCTAATCATAGGATAGAATC
43
72
CTAATTGGCTACGTCATAGAGAGATTCTTTAGGATGAGAAA
42
B7
TACGCGGGCTGAAACCAGGTTTCTTACAATAGGATATGGAG
42
57
TTGAACGGCTGGAGGGATACACCTGGTCATAGGATACCATC
42
66
GCACCCGGCTAGACATAAACTGAGTGCAATAGGATATTTAA
43
A2
AGTGGAGGCTACCTCACAATGGTGGAAATTAGGATGCAACC
43
B5
ATAAAAGGCTCGTTATAGCAAATAACTATTAGGATCATCCA
43
D3
CCACGAGGCTCTGACGATTTCTTTAGGATGGCACG
35
A8
TATAGAGGCTGTCAGATCAATGTGAGGGCTAGGATCTTCAA
43
C2
GTAAAAGGCTAAGACTGAAGGCGAGTAGCTAGGATGCTCTA
41
ermE
GCTAGCCGCGGTCGATCTTGACGGCTGGCGAGAGGTGCGGGGAGGATCTGACCGACGCGG|TCCACACGTGGCACCGCGATGCTGTTGTGGGCACAATCGTGCCGGTTGGTAGGATCCAGCG
122
A1
AGGCCGGGCTGCACTTATTCATCTGTAGATAGGATAACCAA
42
B6
TCAAATGGCTAGGATATAACCTACACGATTAGGATAGCCCC
43
A6
GAGGGAGGCTATGTGATCGGGGAAAGAAATAGGATAGCGGT
42
B10
CTAGGGGGCTATGATAAAATGTGAGTCAATAGGATCGGTAA
42
C4
CTTCAGGGCTTTTGTTTGGCTTAAATCCGTAGGATAAAAAG
41
D10
TAGACTGGCTCGTTGCTAGCGTAACCAAATAGGATACAACG
41
C10
TCTCGCGGCTCTATCGCTTAGGTGGGTAGTAGGATGGCAAG
41
A4
AAAAGCGGCTATATCGCTACAGCTTCCAGTAGGATACAAGA
42
62
CGCCGCGGCTAAAAAAGAAATAGGCTCAATAGGATGAACCC
43
39
CACGACGGCTAAACCAGAAAGGCCTGGCCTAGGATTAATGT
43
B13
TGCTCGGGCTTTGCGGAATTCTAACTACGTAGGATTAGCTT
43
A3
TAGCAGGGCTCCAAAACTAACGCCTGATGTAGGATCAGATG
42
81
CAGGTCGGCTGGTTGGCTGACAAAGTGCGTAGGATGACTGT
43
D4
CCCGTCGGCTGGAAACACTTGATCCAACTTAGGATACTTCC
41
69
TTACCAGGCTCCATAAGTCAAGACGCTTGTAGGATAAAAGT
43
C9
TAAAGGGGCTCCCTCAAAAAAGACTTAGTTAGGATGCCAAG
41
67
TAAAGGGGCTGCCGTATGGTTAAGGTGAGTAGGATCGGAAG
43
65
AGATCTGGCTCCTTTCACAAACGTACTCATAGGATAGGTTT
42
61
CACGCTGGCTACATTATACCTCGGCATAGTAGGATATCCGC
43
A9
CCGGGCGGCTTCCTCATGCTTGACTTGACTAGGATAAAGGG
42
63
CCCCCCGGCTTTTACCATTCGGCAAGTTCTAGGATCCCCAA
42
70
ACCCGTGGCTACTCAGCGAATAACTCTTGTAGGATACATAA
42
49
TGTTTCGGCTCAGTTCTAGACCTACTCATTAGGATTTTTCA
41
74
GGCTCAGGCTGAATGGTCTACCAAGCTATTAGGATCAGCAG
42
21
TGTGCGGGCTCTAACACGTCCTAGTATGGTAGGATGAGCAA
41
D6
TAACGAGGCTTGGAACATTATGGACTCCATAGGATACGTAA
41
tcp
CTGTTGGCTACTCTATACATTGATAGTGGTAGGATCCCTATCAGTGATAGGAGAT
55
71
GTGTATGGCTTCAACGTGGAATACCTCCATAGGATAACGAG
43
ermEp1*
GTTGTGGGCACAATCGTGCCGGTTGGTAGGATCCAGCG
42
B4
GCTGTAGGCTGTTAATATATTTCGGTGTGTAGGATACGGGC
43
C3
CTGAAATGATGTGGAGCACTTTAGGATAAATGA
33
64
ACCTAAGGCTAAACCATTATATAGACTCATAGGATAGCGAG
43
tcp induced
CTGTTGGCTACTCTATACATTGATAGTGGTAGGATCCCTATCAGTGATAGGAGAT
55
68
CAGCCGGGCTCTTATCTTAAACTGGAGCCTAGGATCTCATT
43
C5
TCCGATGGCTGGGGTAAAGACAACCGGGTTAGGATGGTTGT
41
B1
CGTCAGGGCTACTCTGGCAACCAAGCGATTAGGATTGAAGG
42
A5
TCAAGAGGCTCCCTCATTAGAGTCTTATATAGGATGGGCAT
43
C7
TCAAAGGGCTAGTATGTGACTCCCCCATATAGGATACACCT
41
D8
ACAGAAGGCTCCCCGTTGCGCCACATATGTAGGATCGACTG
41

References

1. Theresa Siegl; Bogdan Tokovenko; Maksym Myronovskyi; Andriy Luzhetskyy;  (2013). "Design, construction and characterisation of a synthetic promoter library for fine-tuned gene expression in actinomycetes."  pubmed