Difference between revisions of "Part:BBa K1033282"

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The amount of fluorescence provided by the qRT-PCR with the D-NY15 primers rose after 8 cycles whereas the negative control (pPICZalpha only) started to be amplified at over 29 cycles (non specific amplification). This means that the D-NY15 encoding gene is well expressed in <i>Pichia pastoris</i> with pGAP as a promoter. This promoter is therefore very efficient in the <i>Pichia pastoris</i> background.
 
The amount of fluorescence provided by the qRT-PCR with the D-NY15 primers rose after 8 cycles whereas the negative control (pPICZalpha only) started to be amplified at over 29 cycles (non specific amplification). This means that the D-NY15 encoding gene is well expressed in <i>Pichia pastoris</i> with pGAP as a promoter. This promoter is therefore very efficient in the <i>Pichia pastoris</i> background.

Revision as of 11:15, 27 October 2017

amilCP blue chromoprotein with strong promoter

This is a blue/purple chromoprotein (amilCP: BBa_K592009) with the RBS B0034 (BBa_B0034) and a strong promoter (CP29: BBa_K1033222). The construct has been shown to work in E. coli and should work in Lactobacillus.

(Construct name: pSB1C3-CP29-B0034-amilCP)

See http://2013.igem.org/Team:Uppsala/reporter-genes for more information.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Team KUAS_Korea 2017: characterization

We tested the functionality of K1033222 as a constitutive promoter and expression of amilCP chromoprotein in the Escherichia coli DH5alpha.

DH5alpha containing the control(pSB1C3) and BBa_K1033282, respectively, were cultured in LB medium containing chloramphenicol.

Absorbances were measured every 30 minutes using BioRad SmartSpecPlus at 588 nm and 800 nm.


T--KUAS Korea--amilCP.png
T--KUAS Korea--amilCP.png
T--KUAS Korea--amilCP.png

The amount of fluorescence provided by the qRT-PCR with the D-NY15 primers rose after 8 cycles whereas the negative control (pPICZalpha only) started to be amplified at over 29 cycles (non specific amplification). This means that the D-NY15 encoding gene is well expressed in Pichia pastoris with pGAP as a promoter. This promoter is therefore very efficient in the Pichia pastoris background.