Difference between revisions of "Part:BBa K317024"
 
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It shows the ''E. coli'', which harbors the plasmid encodes this device, the OD660 decreased. Meanwhile, the control didn’t show remarkable decrease. This means ''E. coli'' aggregates from the expression of Antigen 43.
 
It shows the ''E. coli'', which harbors the plasmid encodes this device, the OD660 decreased. Meanwhile, the control didn’t show remarkable decrease. This means ''E. coli'' aggregates from the expression of Antigen 43.
  
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Also we confirmed the expression of Antigen 43 by SDS-PAGE (Fig. 6). Antigen 43 is composed by two subunits, α and β, which the molecular weight are 50 kDa and 53 kDa, respectively. The band was observed in the expected positions (Antigen 43 +). This band was not observed in the sample that doesn’t express Antigen 43  (Antigen 43 -). We can say the decrease of OD660 (described above) comes from the function of Antigen 43.
 
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[[Image:SDS-PAGE.jpg]]
 
  
 
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Latest revision as of 04:00, 28 October 2010

Constitutive promoter(BBa_J23102)-RBS-Antigen 43-Double terminator

Antigen 43 involves in auto-aggregation of the cell. By expressing Antigen 43, the cell will aggregate when the culture solution is stood. Antigen 43 is derived from E. coli genome. The wildtype of Antigen 43 contains 6 PstI sites but this part does not contain any of them. So, this parts can be used as a normal BioBrick which is able to ligate 2 parts by 3A assembly.

In this device, Antigen 43 is expressed constitutively under a strong promoter. The result of measuring OD660 approximately 1 cm from the top of the culture solution is shown below (Fig).

High.jpg

It shows the E. coli, which harbors the plasmid encodes this device, the OD660 decreased. Meanwhile, the control didn’t show remarkable decrease. This means E. coli aggregates from the expression of Antigen 43.


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 770
    Illegal NgoMIV site found at 1994
    Illegal NgoMIV site found at 2504
    Illegal NgoMIV site found at 2525
    Illegal AgeI site found at 1520
    Illegal AgeI site found at 2264
    Illegal AgeI site found at 2678
    Illegal AgeI site found at 2920
  • 1000
    COMPATIBLE WITH RFC[1000]