Difference between revisions of "Part:BBa K883151"

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<partinfo>BBa_K883151 parameters</partinfo>
 
<partinfo>BBa_K883151 parameters</partinfo>
  
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Temperature Stability
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Like the wild type CCMV, the {} is stable at 4 degrees Celcius.
  
 
pH stability
 
pH stability
  
 
Time stability
 
Time stability

Revision as of 11:21, 24 September 2012

CCMV coat protein gene 25AA N-terminal replaced with HIStag under IPTG inducable promotor

This part produces a coat protein of the Cowpea Clorotic Mottle Virus (CCMV), with the first 25 amino acids replaced with an Histidine tag (6x).

Usage and Biology

CCMV is an abriviation of the Cowpea Chlorotic Mottle Virus, one of the most studied viruses in producing Virus Like Particles (VLPs). This part is the produces a coat protein of the Cowpea Clorotic Mottle Virus (CCMV), with the first 25 amino acids replaced with an Histidine tag (6x).When expressed in Escherichia coli, these coat proteins can be harvested and be assembled in vitro using the Wageningen_UR iGEM 2012 protocols.


Protocols for production and purification:

  • [http://2012.igem.org/Team:Wageningen_UR/Protocol/StartupCCMV| CCMV production protocol part 1]
  • [http://2012.igem.org/Team:Wageningen_UR/Protocol/DialysisCCMV| CCMV production protocol part 2]
  • [http://2012.igem.org/Team:Wageningen_UR/Protocol/RoundupCCMV| CCMV purification protocol]

[ADD PICTURE EM]

This part is made from the wild type CCMV coat protein. CCMV is the abbreviation of Cowpea Chlorotic Mottle Virus, and is a well studied black eyed pea infecting virus. It is a virus in the family Bromoviridae, and it is one of the most studied plant viruses. As from the discovery of the virus in 1967, it was an interesting virus due to the easy cultivation and harvesting procedures involved. Interest in this virus is especially in the ability to disassemble the virus, remove the content and refold the empty particle simply by altering the pH. This makes this virus one of the first and best studied Virus Like Particles.

Besides having the characteristics of the wild type CCMV coat protein, a HIStag is added to it. This means purification with a nickel column is possible to use to purify the VLPs. (Untested)


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]

Functional Parameters

Like the wild type CCMV, the {} is stable at 4 degrees Celcius.

pH stability

Time stability