Difference between revisions of "Part:BBa K3598042"

 
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This part is a tightly repressible induction system,  PJExD promoter is regulated by EilR, and  several cationic dyes act as efficient low-cost inducers. sfGFP is used for quantitative analysis of the system.
 
This part is a tightly repressible induction system,  PJExD promoter is regulated by EilR, and  several cationic dyes act as efficient low-cost inducers. sfGFP is used for quantitative analysis of the system.
  
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===Usage and Biology===
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===Overall design of the part===
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To achieve co-expression of adhesive/cohesive protein and tyrosinase, we need a new inducible system different from T7-LacI inducible promoter to control the expression of tyrosinase. Thus, we constructed BBa_K3598042, which consists EilR repressor and PJExD promoter and uses cationic dye as inducer.
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The EilR promoter promotes the expression of the EilR, the product of which suppresses the PJExD promoter, thus preventing the expression of the sfGFP. We verified the properties of the cationic dye induced expression system formed by the composite part under the influence of 7 different dyes, by transforming it into E. Coli BL21 and examining the expression of the sfGFP. When cationic dyes are added, the suppression of EilR's expression on the PJExD promoter is eliminated, therefore allowing for the expression of the sfGFP sequence, which makes the cells florescent.
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[[File:T--BEIJING_4ELEVEN--New_Composite_Part_Figure_1.png|400px|thumb|center|Figure 1 The overall design of BBa_K3598042]]
  
 
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Revision as of 06:15, 26 October 2020


EilR_EilR promoter_PJExD promoter_sfGFP

This part is a tightly repressible induction system, PJExD promoter is regulated by EilR, and several cationic dyes act as efficient low-cost inducers. sfGFP is used for quantitative analysis of the system.


Overall design of the part

To achieve co-expression of adhesive/cohesive protein and tyrosinase, we need a new inducible system different from T7-LacI inducible promoter to control the expression of tyrosinase. Thus, we constructed BBa_K3598042, which consists EilR repressor and PJExD promoter and uses cationic dye as inducer.

The EilR promoter promotes the expression of the EilR, the product of which suppresses the PJExD promoter, thus preventing the expression of the sfGFP. We verified the properties of the cationic dye induced expression system formed by the composite part under the influence of 7 different dyes, by transforming it into E. Coli BL21 and examining the expression of the sfGFP. When cationic dyes are added, the suppression of EilR's expression on the PJExD promoter is eliminated, therefore allowing for the expression of the sfGFP sequence, which makes the cells florescent.

Figure 1 The overall design of BBa_K3598042

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 916
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 346
    Illegal AgeI site found at 496
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 954