Difference between revisions of "Part:BBa K592009:Experience"

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<I>iGEM 2013 Hong_Kong_HKUST Trevor Y.H. HO</I>
 
<I>iGEM 2013 Hong_Kong_HKUST Trevor Y.H. HO</I>
 
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This part was employed in testing our Gibson Assembly protocol. In the experiment design, the mRFP coding DNA sequence in J04450 was replaced with that of amilCP.  
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This part was employed in testing our Gibson Assembly protocol. In the experiment design, the mRFP coding DNA sequence in J04450 was replaced with that of amilCP. The resulting plate from transformation of the post-reaction mixture is shown. The color of the blue chromoprotein was clearly distinguished from that of mRFP, which aided in my evaluation of the performance in Gibson Assembly.
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[[Image:GA06_01Aug2013_Expt3.jpg]]
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An image showing a streaked plate of DH10b carrying pSB1C3-R0010-B0034-K592009-B0015.
 
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Revision as of 14:17, 3 August 2013

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_K592009

User Reviews

UNIQff0eecda122a5d05-partinfo-00000000-QINU UNIQff0eecda122a5d05-partinfo-00000001-QINU

No review score entered. iGEM 2013 Hong_Kong_HKUST Trevor Y.H. HO

GA06 01Aug2013 Expt3.jpg This part was employed in testing our Gibson Assembly protocol. In the experiment design, the mRFP coding DNA sequence in J04450 was replaced with that of amilCP. The resulting plate from transformation of the post-reaction mixture is shown. The color of the blue chromoprotein was clearly distinguished from that of mRFP, which aided in my evaluation of the performance in Gibson Assembly. GA06 01Aug2013 Expt3.jpg An image showing a streaked plate of DH10b carrying pSB1C3-R0010-B0034-K592009-B0015.

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BBa_K592009 iGEM Groningen 2012

Our team has managed to couple this biobrick part with our promoters: alsT, fnr, and sboA. The cloning was done in BBa_K818000 (plasmid backbone for B. subtilis, engineered by team Groningen 2012), to allow color expression in B. subtilis. We utilized a strong RBS BBa_B0034 for pigment expression in E. coli and B. subtilis.

The purple/blue colour was strongly visible in E.coli without any induction, while the expression in B. subtilis was more subtle (B. subtilis colony looks slightly blue on the plate agar).

After induction of the promoter before AmilCP, B. subtilis also turned clearly purple/blue. Please have a look at the page of BBa_K818400 for more information.



No review score entered. User:agynna

iGEM Team Uppsala University 2012

We have observed that expression of amilCP confers a noticeable fitness cost in E coli. This is surprising, given that such behaviour is not seen in other homologous chromoproteins and fluorescent proteins. We thus recommend using the new aeBlue (BBa_K864401) chromoprotein for blue color expression, which also has a more clear blue color.

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