bacteriophage PL-λ promoter

来源: E. coli K2 MG655
功能: promoter

Summary

基本信息

定量方法 1. Approximately 20 μl of overnight cultures of library clones growing in LB broth were used to inoculate into 5 ml of M9G medium and the cultures were grown at 37°C with orbital shaking. After 14 h, the centrifuged cells were resuspended in ice-cold water. Flow cytometry was performed on a Becton Dickinson FACScan, and the geometric mean of the fluorescence distribution of each clonal population was calculated. 2. Transcriptional analysis: Cultures inoculated as previously were grown for 3 h, and the total RNA was extracted and subjected to quantitative RT-PCR. 3. Chloramphenicol resistance: various concentrations of chloramphenicol ( 0-500 μg/ml) was used to test the promoter strength. After overnight incubation at 37°C, the lowest concentration of chloramphenicol that inhibited the growth of a clone was recorded. 4. Promoter strength metric calculated using a previously published dynamic model (Leveau, J., et al. (2001)J. bacteriol., 183, 6752-6762.).
培养条件 Assay strains were grown at 37℃with 225 rpm orbital shaking in M9-minimal media (Maniatis, T., et al. Molecular Cloning: A Laboratory Mannual. (Cold Spring Harbor Lab. Press, Plainview, NY)) containing 5 g liter D-glucose. When necessary, the M9 media were supplemented with 0.1% casamino acids.
报告基因 GFP
诱导剂 IPTG
提交人 Xing Yan;Wenfang Zhao

元件序列

元件名 序列 序列长度
a mutated bacteriophage PL-λ promoter (N)
CAATTCCGACGTCTAAGAAACCATTATTATCATGACATTAACCTATAAAAATAGGCGTATCACGAGGCCCTTTCGTCTTCACCTCGAGTCCCTATCAGTGATAGAGATTGACACCCCTATCAGTGCTAGAGATACTGAGCACATCAGCAGGACGCACTGACC
162
a mutated bacteriophage PL-λ promoter (II)
CAATCCGACGTCTAAGAAACCATTATTATCATGACATTAACCTATAAAAATAGGCGTATCACGAGGCCCTTTCGTCTTCACCTCGAGTCCCTATCAGTGATAGAGATGGACATCCCTATCAGTGATAGAGATACTGAGCACATCAGCAGGACGCACTGACC
161
a mutated bacteriophage PL-λ promoter (OO)
CAATTCCGACGTCTAAGGAAACCATTATCATGACATCAACCTATAAAAATAGGCGTATCACGAGGCCCTCTCGTCTCCACCTCAAGCTCCCTATCTAGTGATAGCGATTGACATCCCTATCAGTGACGGAGATATTGAGCACATCAGCAGGACGCACTGACC
162
a mutated bacteriophage PL-λ promoter (U)
CAATTCCGACGTCTAAGAAACCATTATTATCATGACACTAACCTATAAAAATAGGCGCATCACGAGGCCCTTTCGTCTCCACCTCAAGCTCCCTATCAGTGATAGAGATTGACATCCCCGCCGGTGATAGAGACACTGAGCACATCAGCAGGACGCACTGACC
163
a mutated bacteriophage PL-λ promoter (K)
CAATTCCGACGTCTAAGAAACCATTATTATCATGACATTAACCTATAAAAATAGGCGTATCACGAGGCCCTCTCGTCTTCACCTCGAGTCCCTATCAGTGATAGGGATTGACATCCCTATCAGTGATAGAGACACTGGGCACATCAGCAGGACGCACTGACC
162
a mutated bacteriophage PL-λ promoter (V)
CAATTCCGACGTCTGAGAGGCCATTATTATCGTGGCATTGGCCTATAAAGGCAGGCGTGTCACGAGACCCTCTCGTCTCCGCCTCGGGTCCCTATCAATGGTAGAGATTGACATCCCCATCAGTGGTGGAGATACTGAGCACATCAGCAGGACGCACTGACC
162
a mutated bacteriophage PL-λ promoter (AA)
CAATTCCGACGTCTAAGAAACCATTATTATTATGACATTAACCTATAAAAATAGGCGTATCACGAGGCCCTTCCGTCTTCACCTCGAGTCCCTATCAGTGATAGAGATTGACATCCCTATCAGTGATAGAAATACTGAGCACATCAGCAGGACGCACTGACC
162
a mutated bacteriophage PL-λ promoter (T)
CAATTCCGACGTCTAAGAAGCCATTACTATCATGACATTAACCTATAGGAATAGGCGTATCACGGGGCCCTTCCGCCTTCACCTCGGATCCCTGTCAGTGCTAGAGATTGACATCCCTACCGGTGATAAAGATACTGAGCACATCAGCAGGACGCACTGACC
162
a mutated bacteriophage PL-λ promoter (B)
CAATTCCGACGTCTAAGAAGCCATTATTATCATGACATTAACCTATAAAAGTAGGCGTGTCACAAGGCCCTTTCGTCTTCACCTCGAGTCCCTATCAGCGATAGAGATTGACATCCCTATCAGTGACCGAGATACTGAGCACATCAGCAGGACGCACTGACC
162
a mutated bacteriophage PL-λ promoter (E)
CAATTCCGACGCCTAAGAAACCATTATTATCATGACATTAGCCTATAAAAATAGGCGTACCACGAGGCCCTTTCGTCTTCACCTCGAGTCCCTATCAGTGATAGAGATTGACACCCCTATCAGTGATAGAGATACTGAGCACATCAGCAGGACGCACTGACC
162
a mutated bacteriophage PL-λ promoter (JJ)
CAATTCCGACGTCTAAGAAACCATTATTATCATGACATTAACCTATAAAAATAGGCGTATCACGAGGCCCTTTCGTCTTCACCTCGAGTCCCTATCAGTGATAGAGATTGACCTCCCTATCAGTGATAGAGATACTGAGCACATCAGCAGGACGCACTGACC
162
a mutated bacteriophage PL-λ promoter (Y)
CAATTCCGACGTCTAAGAAACCATTATTATCATGACATTAACCTATAAAAATAGGCGTATCACGAGGCCCTTTCGTCTTCACCTCGAGTCCCTATCAGTGATAGGGATTGACATCCCTATCAGTGATAGAGACACTGAGCACATCAGCAGGACGCACTGACC
162
a mutated bacteriophage PL-λ promoter (P)
CAATTCCGACGTCTAGGAAACCGTTATATCATGACGCCGACCTATAAAGATAGGCGCGTCNCGAGGCCCTTCCGCCTTCACCTCGNGCTCCCTATCAGTAATAGAGATTGACACCCCTGTCAGTGATAGGGATACTGAGCACATCAGCAGGACGCACTGACC
162
a mutated bacteriophage PL-λ promoter (S)
CAATTCCGACGTCTAAGAAACCATTATTATCATGACATTAACCTATAAAAATAGGCGTATCACGAGGCCCTTTCGTCTTCACCTCGAGTCCCTATCAGTGATAGAGATTGACATCCCTATCAGTGATAGAGACACTGAGCACATCAGCAGGACGCACTGACC
162
a mutated bacteriophage PL-λ promoter (L)
CAATTCCGACGTCTAAGAAACCATTGTTATCATGACATTAACCTATAAAAATAGGCGTATCACGAGGCCCTTTCGTCTTCACCTCGAGTCCCTATCAGTGATAGAGATTGACACCCCTATCAGTGACAGAGATACTGAGCACATCAGCAGGACGCACTGACC
162
a mutated bacteriophage PL-λ promoter (R)
CAATTCCGACGTCTGAGAAGCCATTATTATCATGACATTAACCTATAAAAGTAGGCGTATCACGAGGCCCTTTCGTCTTCACCTCAAGCCCCTATCAGTGATAGAGATTGACATCCCTATCAGTGATAGAGACACTGAGCACATCAGCAGGACGCACTGACC
162
a mutated bacteriophage PL-λ promoter (Q)
CAATTCCGACGTCTAAGAAACCATTATTATCATGACATTAACCTATAAAAATAGGCGTATCACGAGGCCCTTTCGTCTTCACCTCGAGTCCCTATCAGTGATAGAGATTGACATCCCTATCAGTGATAGGGATACTGAGCACATCAGCAGGACGCACTGACC
162
the wild-type bacteriophage PL-λ promoter (Lutz, R. & Bujard, H. (1997) Nucleic Acids Res. 25, 1203鈥?210.)
CAATTCCGACGTCTAAGAAACCATTATTATCATGACATTAACCTATAAAAATAGGCGTATCACGAGGCCCTTTCGTCTTCACCTCGAGTCCCTATCAGTGATAGAGATTGACATCCCTATCAGTGATAGAGATACTGAGCACATCAGCAGGACGCACTGACC
162
a mutated bacteriophage PL-λ promoter (Z)
CAATTCCGACGTCTAAGAAACCATTATTATCATGACATTAGCCTATAAAAATAGGCGTATCACGAGGCCCTTTCGCCTTCACCTCGAGTCCCTATCGGTGACAGAGGTTGACATCCCTATCGGTGATAGAGATACTGAGCACATCAGCAGGACGCACTGACC
162
a mutated bacteriophage PL-λ promoter (W)
CAATTCCGACGTCTAAGAAACCATTATTATCATGACATTAACCTATAAAAATAGGCGTATCACGAGGCCCTTTCGTCTTCACCTCGAGTCCCTATCAGTGATAGAGATTGACATCCCTATCAGTGATAGAGATACTGAGCACATCAGCAGGACGCACTGACC
162
a mutated bacteriophage PL-λ promoter (BB)
CAATTCCGACGTCTAAGAGACCATTATTATCGTGACATTAACCTATAAGAACAGGCGTGTCACGAGGCCCTTTCGTCTTCACCTCGAGTCCCTATCAGTGACAGAGATTGACACCCCTATCAGTGATAGAGATACTGAGCACATCAGCAGGACGCACTGACC
162
a mutated bacteriophage PL-λ promoter (F)
CAATTCCGACGTCTAAGAAACCATTGTTATCGTGACATTAACCTATAANANTAGGCGTATCACGAGGCCCTTTCGCCTTCACCTCGAGTCCCTATCAGYGATAGAGACCGACACCCCTATCAGTGATAGGGATACTGGGCACATCAGCAGGACGCACTGACC
162

References

1. Alper H,Fischer C,Nevoigt E,Stephanopoulos G (2005). "Tuning genetic control through promoter engineering."  Proc Natl Acad Sci U S A 102(36);12678-83