RBS of Streptomyces from PNAS-2015-Bai

来源: Streptomyces venezuelae ISP5230
功能: RBS

Summary

native RBSs and sequenced synthetic RBSs

基本信息

定量方法 Protoplasts was prepared from mycelia of Streptomyces at first and then the protoplasts were analyzed by BD FACSCalibur Flow Cytometer with a 488-nmexcitation laser and the FL1 (530/30 nm band-pass filter) detector. Eachsample collected 50,000 events, and the data were further acquired using BDFACSuite software and analyzed by FlowJo 9.3.2 software (Tree Star, Inc.). The parameters of the FACS setting were as follows: FSC-E00, SSC-650, FL1-400, FL2-400; threshold: FSC-50, SSC-400. All date compared with the date of strong promoter kasOp*
培养条件 Mannitol soya flour medium was used for conjugation, and malt extract–yeast extract–maltose medium was used for liquid inoculation of the spores of S. venezuelae ISP5230. The media were supplemented with different antibiotics at appropriate concentrations as follows: ampicillin at 100 μg/mL, apramycin at 50 μg/mL, nalidixic acid at 25 μg/mL, kanamycin at 25 μg/mL, and chloramphenicol at 25 μg/mL Spores of S. avermitilis WT transformants were used to inoculate a 250-mL flask containing 25 mL seed medium [glucose (5 g), soy flour (15 g), and yeast extract (5 g) per liter, at pH 7.2], and the culture was allowed to grow with shaking (220 rpm) at 30 °C for 2 d. A 1-mL aliquot of the culture was used to inoculate a 250-mL flask containing 50 mL production medium, CaCO3 (5 g), and yeast extract (2 g) per liter at pH 7.0] (41). The samples were incubated with shaking (220 rpm) at 28 °C for 5 d.
报告基因 superfolder green fluorescent protein
诱导剂
提交人 Yan Xing;Zhao Wenfang

元件序列

元件名 序列 序列长度
SR12
cgcgcctaaggagtgtccatA
21
R3
agattttttcgggggtgaccA
21
R1
cactcgcagaggagattgcaA
21
SR25
tctaagtaaggagcggtctaA
21
SR18
tctcctcaaggagtgtccatA
21
R7
tcatgacgtaagggggcgctA
21
SR27
tctaagtaaggagttgaataA
21
R4
gtcgagcgaagggaacggcaA
21
SR3
ctggatgttaaagtgtccatA
21
SR24
gcgtgtgaaggagtgtccatA
21
SR31
tgccgctaaggagtgtccatA
21
SR6
aaagcatctagagtgtccatA
21
SR20
tctaagtaaggaggccgcacA
21
SR11
tcagacgtcggagtgtccatA
21
SR33
tctaagtaaggagttcagccA
21
R10
cgacgacctaggggttttgcA
21
SR5
tctaagtaagagtgttttccA
21
R13
gtatctgaaaggggatacgcA
21
SR38
tctaagtaaggagtagcgatA
21
R5
agtaaggaacctgcgcttctA
21
SR21
tctagtaaggagggaagagA
20
SR34
acatacaaaggagtgtccatA
21
R15
tctaagtaaggagtgtccatA
21
SR15
tctaagtaaggagggtacgaA
21
SR2
tctaagtaagtctacaatacA
21
SR37
tctaagtaaggagacatctaA
21
R11
cacaaggggttgtgaccgggA
21
SR10
tctaagtaagccgggtaaaaG
21
SR39
ttcgagtaaggagtgtccatA
21
SR16
tctaagtaaggaggtcaacaA
21
SR4
gaacttaaaggagtgtccatA
21
SR30
tctaagtaaggagagacataA
21
R2
ttctcgcgggtgctgcctggA
21
SR1
tctaagtaagcaggactgtaT
21
R14
cctaacgaggagatcggttcA
21
SR40
cttgtcaaaggagtgtccatA
21
SR41
tctaagtaaggagtaggctgA
21
R8
cgtgcaggactgggggagttA
21
R6
tgagtgaagggagttgccacA
21
SR13
gaaaaaaaaggagtgtccatA
21
SR9
caaaaatcgtgagtgtccatA
21
SR26
cacctaaaaggagtgtccatA
21
SR22
ccgccccaaggagtgtccatA
21
R9
aactacgaaggggagtcagtA
21
SR8
tctaagtaagcgaactattcA
21
SR23
tctaagtaaggattacttgcA
21
R12
caccgacagacggagacgtaA
21
SR36
tctaagtaaggagagccggtA
21
SR32
tctaagtaaggagactgcacA
21
SR19
tctaagtaaggagaataagtA
21
SR35
tagccttaaggagtgtccatA
21
SR28
catgccattggagtgtccatA
21
SR14
aaacgggaaggagtgtccatA
21
SR17
tctaagtaaggagcttataaA
21
SR29
tctaagtaaggagtaaccgcA
21
SR7
gctacccgtagagtgtccatA
21

References

1. Chaoxian Bai; Yang Zhang; Xuejin Zhao; Yiling Hu; Sihai Xiang; Jin Miao; Chunbo Lou; Lixin Zhang;  (2015). "Exploiting a precise design of universal synthetic modular regulatory elements to unlock the microbial natural products in Streptomyces."  pubmed