Difference between revisions of "Part:BBa K292006:Experience"

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<partinfo>BBa_K292006 AddReview 1</partinfo>
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<I>NTNU_Trondheim 2012</I>
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We decided to improve this part since last years team had problems with it. We ordered the biobrick from the registry, test cut it, and sequenced it. In addition we reassembled the the parts BBa_B0030, BBa_C0012 and BBa_B0014, test cut these, and sent them to sequencing. The results can be seen in the photo below.

Revision as of 20:36, 26 September 2012

This experience page is provided so that any user may enter their experience using this part.
Please enter how you used this part and how it worked out.

Applications of BBa_K292006

Reliable to use when we want to control a gene expression by using pLac promoter. In fact when LacI is not expressed, pLac is activated, and when there is an expression of LacI, pLac is inhibited. We can easily design a system to induce the transcription of LacI by using another operon as the tetracycline operon and creates a failed feedback mechanism for pLac inhibition and so, for gene expression inhibition.


User Reviews

UNIQ6d8d5f4b056a7061-partinfo-00000000-QINU



•

NTNU_Trondheim 2011

We found and orderd this part from the parts registry. Since it had RBS and double terminator it would save us from some cloning work.

When testcutting the constructs containing BBa_K292006 we got some unexpected results. Therefore we decided to digest the brick with three differnt enzymes with restriction sites in the given biobrick-sequence in addition to EcoR1 and PstI (for looking at the biobrick length) Except from the sample cut with EcorR1 and PstI, the samples were also cut with BglI wich has a recognition site in the pSB1A2 backbone.

LacI2 plasmid cut with EcoR1 + PstI, BglI + BclI, BglI + EcoRV, BglI + BanII

Overwiev of restrctions and expected fragmentlengths:

Restriction enzymes Expected lengths (bp)
EcoR1 + PstI 2053 (backbone) and 1303 (biobrick)
BglI + BclI 2032 + 1324
BglI + EcoRV 1760 + 1596
BglI + BanII 1835 + 1521

As we can see on the resulting gel none of the restriction sites inn the LacI2 fragment appear to be present. In additon the LacI2 insert looks smaller than it's supposed to be. This indicates that the biobrick sequence does not match the sequence given in the parts registry.



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UNIQ6d8d5f4b056a7061-partinfo-00000002-QINU


UNIQ6d8d5f4b056a7061-partinfo-00000003-QINU



•

NTNU_Trondheim 2012

We decided to improve this part since last years team had problems with it. We ordered the biobrick from the registry, test cut it, and sequenced it. In addition we reassembled the the parts BBa_B0030, BBa_C0012 and BBa_B0014, test cut these, and sent them to sequencing. The results can be seen in the photo below.