Difference between revisions of "Part:BBa K1489000:Experience"

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This experience page is provided so that any user may enter their experience using this part.<BR>Please enter
 
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<I>Iowis Zhu</I>
 
<I>Iowis Zhu</I>
 
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Expression tests
 
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BBa_K148900 was transformed into chemically competent DH5-alpha cells and plated overnight with chloramphenicol challenge. One colony was incubated in 5 mL LB media + 34 ug/mL chloramphenicol and grown to mid-log phase at 37oC. Arabinose was added to a final concentration of 0.2% to initiate protein production and cells were re-incubated at 37 oC for 21 hours. Cells were then pelleted and lysed with 10% SDS solution. Cell lysate was centrifuged at 13200 rpm in a tabletop centrifuge for 1 hour to separate soluble and insoluble fractions. Both fractions were loaded on an SDS-PAGE gel for western blot analysis using anti-His antibodies.
 
  
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[[File:BovineGalectinExpression.jpg]]
 
  
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Clear band of correct size can be seen in the soluble induced fraction, while no band is seen in the uninduced cells.
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<partinfo>BBa_K1489000 Addreview 5</partinfo>
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<I>Iowis Zhu</I>
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We have found that this biobrick can be functionally expressed in E. coli. This was confirmed via protein purification followed by quantification of a binding constant.
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[[File:Bovine_Binding_Assay.png]]
 
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Latest revision as of 06:22, 26 October 2014

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Iowis Zhu

We have found that this biobrick can be functionally expressed in E. coli. This was confirmed via protein purification followed by quantification of a binding constant.

Bovine Binding Assay.png UNIQd65b29f0f30892fa-partinfo-00000002-QINU