Difference between revisions of "Part:BBa K660201:Design"
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__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K660201 short</partinfo>
 
<partinfo>BBa_K660201 short</partinfo>
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<h3>Origin</h3>
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Horse sweat <i>(Equus caballus)</i><br/>
  
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<partinfo>BBa_K660201 SequenceAndFeatures</partinfo>
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<h3>Function</h3>
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Latherin is a surfactant protein with detergent like properties. <br/>
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Its normal biological function is temperature regulation and is believed to function by enhancing evaporation from the pelt. It does so by binding to hydrophobic surfaces and allowing them to become wettable.  <Br/>
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<h3>Applications</h3>
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Latherin is able to temporarily bind to hydrophobic surfaces and has therefore been investigated as a possible method of breaking up biofilms. It is also suspected that the protein may have antimicrobial functions.  <br/>
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The 2011 University of Glasgow iGEM team used this part in their system, DISColi, to aid in the dispersal of biofilms.
  
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<h3>Additional Information</h3>
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Note: Latherin is suspected to be the cause of allergic reaction in individuals with an allergy to horses.<br/>
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<h3>Illegal Restriction Sites</h3>
  
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===Design Notes===
 
 
We are aware of two illegal restriction sites that exist in our latherin biobrick.  Due to time constraints, we were unable to complete site directed mutagenesis. The primers we designed for this are below, for any future team.<br/><br/>
 
We are aware of two illegal restriction sites that exist in our latherin biobrick.  Due to time constraints, we were unable to complete site directed mutagenesis. The primers we designed for this are below, for any future team.<br/><br/>
  
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MTP= 78.71<br/><br/>
 
MTP= 78.71<br/><br/>
  
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<h3>References & Further Reading</h3>
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===Source===
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Latherin was originally isolated from horse sweat. It was expressed in <i>E.coli</i> by researchers at University of Glasgow. We received it from them in a plasmid and amplified it up by PCR to add a biobrick prefix and suffix and a 6xHIS tag.
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===References===
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Beegley J, et al., 1986. Isolation and characterization of latherin, a surface-active protein from horse sweat<br/>http://www.ncbi.nlm.nih.gov/sites/entrez?db=pubmed&cmd=search&term=3753435>.  
 
Beegley J, et al., 1986. Isolation and characterization of latherin, a surface-active protein from horse sweat<br/>http://www.ncbi.nlm.nih.gov/sites/entrez?db=pubmed&cmd=search&term=3753435>.  
 
<br/><br/>
 
<br/><br/>
 
Kennedy M., 2011., Latherin and other biocompatible surfactant proteins<br/>
 
Kennedy M., 2011., Latherin and other biocompatible surfactant proteins<br/>
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http://www.biochemsoctrans.org/bst/039/bst0391017.htm<br/><br/>
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http://www.biochemsoctrans.org/bst/039/bst0391017.htm
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Information on "Affinity Purification" <br/>
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<partinfo>BBa_K660201 SequenceAndFeatures</partinfo>
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http://www.brunschwig-ch.com/pdf/downloads/ABT_RapidRun_Procedure.pdf?PHPSESSID=cb79f5862da62a654bb1a6aefe87c62b
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Revision as of 04:06, 22 September 2011

Latherin (Surfactant Protein) + 6xHIS Tag

Origin

Horse sweat (Equus caballus)

Function

Latherin is a surfactant protein with detergent like properties.

Its normal biological function is temperature regulation and is believed to function by enhancing evaporation from the pelt. It does so by binding to hydrophobic surfaces and allowing them to become wettable.

Applications

Latherin is able to temporarily bind to hydrophobic surfaces and has therefore been investigated as a possible method of breaking up biofilms. It is also suspected that the protein may have antimicrobial functions.
The 2011 University of Glasgow iGEM team used this part in their system, DISColi, to aid in the dispersal of biofilms.

Additional Information

Note: Latherin is suspected to be the cause of allergic reaction in individuals with an allergy to horses.

Illegal Restriction Sites

We are aware of two illegal restriction sites that exist in our latherin biobrick. Due to time constraints, we were unable to complete site directed mutagenesis. The primers we designed for this are below, for any future team.

1st PstI site:
5': GCG GAA ATT AAA CTG CAA GAT ACC CGT CTG CAA CTG

    Forward Primer: 5' CAG TTG CAG ACG GGT ATC TTG CAG TTT AAT TTC CGC

    Reverse Primer: 5' GCG GAA ATT AAA CTG CAA GAT ACC CGT CTG CAA CTG

MTP= 79.33


2nd PstI site:
5': GCG ATT GAG CTG CAA AGC GGC AAT CCG C

    Forward Primer: 5' G CGG ATT GCC GCT TTG CAG CTC AAT CGC

    Reverse Primer: 5' GCG ATT GAG CTG CAA AGC GGC AAT CCG C

MTP= 78.71

References & Further Reading

Beegley J, et al., 1986. Isolation and characterization of latherin, a surface-active protein from horse sweat
http://www.ncbi.nlm.nih.gov/sites/entrez?db=pubmed&cmd=search&term=3753435>.

Kennedy M., 2011., Latherin and other biocompatible surfactant proteins
http://www.biochemsoctrans.org/bst/039/bst0391017.htm


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 691
    Illegal PstI site found at 922
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 691
    Illegal PstI site found at 922
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 462
    Illegal BglII site found at 1005
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 691
    Illegal PstI site found at 922
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 691
    Illegal PstI site found at 922
  • 1000
    COMPATIBLE WITH RFC[1000]