Difference between revisions of "Part:BBa K2556333"

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===<h1>References</h1>===
 
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<!-- Uncomment this to enable Functional Parameter display
 
<!-- Uncomment this to enable Functional Parameter display

Revision as of 13:33, 14 October 2018

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Light control system with eGFP


Under dark conditions, the phosphate group is transferred from Yf1 protein to FixJ protein, the phosphorylated FixJ protein activates the PfixK2 promoter and then activates the downstream gene expression. When induced by light, the phosphorylation of Fixj protein will be blocked, and the expression of genes regulated by PfixK2 promoters will be inhibited. T--ZJUT-China--part101.png

Characterize

To further verify the effectiveness of the system, the plasmids with the system were transferred into different hosts and cultured for more than 24 hours under blue light and dark light respectively. The fluorescence value of eGFP in the culture medium was measured.

Experimental Results

Detection of expression efficiency of primary light control system in different hosts

T--ZJUT-China--designyu7.png

Figure 1. expression efficiency in DH5α

T--ZJUT-China--designyu8.png

Figure 2. expression efficiency in BL21

T--ZJUT-China--part3.png

Figure 3. expression efficiency in 4MG1655

T--ZJUT-China--designyu10.png

Figure 4. expression efficiency in BW25113

The dusk-eGFP-pUC57 plasmid was transferred into the host of DH5α,BL21,4MG1655 and BW25113, cultured under dark or blue light for a period of time and the fluorescence value of eGFP was detected. After 24 hours, the fluorescence value of the experimental group in the dark culture showed an obvious increasing trend. There was no significant increase in fluorescence value in both the experimental group under blue light and the negative control group.



Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 605
    Illegal NgoMIV site found at 677
    Illegal NgoMIV site found at 767
    Illegal NgoMIV site found at 785
    Illegal NgoMIV site found at 1297
    Illegal NgoMIV site found at 1590
    Illegal NgoMIV site found at 1684
    Illegal AgeI site found at 319
    Illegal AgeI site found at 1465
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1354
    Illegal BsaI.rc site found at 218

References