Part:BBa_K2556333
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.
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
Figure 2. expression efficiency in BL21
Figure 3. expression efficiency in 4MG1655
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
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE 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 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 1354
Illegal BsaI.rc site found at 218
References
| None |