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Revision as of 18:02, 31 October 2017


LacIq

This part contains LacIq repressor, LacIq spacer, LacIq promotor and LacIq coding sequence and is used for Rhodobacter sphaeroides 2.4.1, cells with the device can carry out inducible expression of the target proteins.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 84
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]



sYFP2(BBa_K2308003) is the key part in our experiment. It is used both for fusion expression (see BBa_K2308002) and inducible cytoplasm expression (combined with BBa_K2308016), and the host cell is Rhodobacter sphaeroides 2.4.1.


Figure 1：PCR test of BBa_K2308003(optimized sYFP2) and original sYFP2



In inducible cytoplasm expression experiment, we constructed the inducible plasmid pIND4 using part BBa_K2308016 and succeeded in turning the Rhodobacter sphaedoides 2.4.1 into inducible strains.IPTG was added （800 μM in final volume）when the OD700 of the strain was about 0.4（grown for about 24h）.

Figure 2：Confirmed map of plasmid pIND4

Figure 3：fluorescent image of sYFP2 (original) and sYFP2(optimized)



Figure 4：Fluorescence intensity of WT, sYFP2(optimized), sYFP2(unoptimized) strains.



It is obvious that after the codon optimization, sYFP2 can be better expressed in Rhodobacter sphaeroides 2.4.1, and the LacIq promotor also worked very well.