Difference between revisions of "Part:BBa K404006"
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<partinfo>BBa_K404006 short</partinfo>
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<partinfo>BBa_K404007 short</partinfo>
  
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{| style="color:black; margin: 0px 0px 500px 20px;" cellpadding="6" cellspacing="1" border="2" align="right"
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! colspan="2" style="background:#66bbff;"|[https://parts.igem.org/Part:BBa_K404007 AAV2-VP123]
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|-
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! colspan="2"|[[image:Freiburg10 p5 promoter.png|200px]]
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|-
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|'''BioBrick Nr.'''
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|[https://parts.igem.org/Part:BBa_K404007 BBa_K404007]
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|-
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|'''RFC standard'''
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|[https://parts.igem.org/Help:Assembly_standard_25 RFC 25]
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|-
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|'''Requirement'''
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|pSB1C3<br>
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|-
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|'''Source'''
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|pAAV_RC from Stratagene
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|-
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|'''Submitted by'''
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|[http://2010.igem.org/Team:Freiburg_Bioware FreiGEM 2010]
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|}
 
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The AAV capsid consists of 60 capsid protein subunits. The three cap proteins VP1, VP2, and VP3 are encoded in an overlapping reading frame. Arranged in a stoichiometric ratio of 1:1:10, they form an icosahedral symmetry. The mRNA encoding for the cap proteins is transcribed from p40 and alternative spliced to minor and major products. Alternative splicing and translation initiation of VP2 at a nonconventional ACG initiation codon promote the expression of VP1, VP2 and VP3. The VP proteins share a common C terminus and stop codon, but begin with a different start codon. The N termini of VP1 and VP2 play important roles in infection and contain motifs that are highly homologous to a phospholipase A2 (PLA2) domain and nuclear localization signals (BR)(+). (Quelle)
  
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<h3>Characterization</h3>
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<h3>References</h3>
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<h1>Usage and Biology</h1>
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<b>Cassinotti, P., Weitzand, M., & Tratschin, J. D.</b>, 1988. Organization of the adeno-associated virus (AAV) capsid gene: mapping of a minor spliced mRNA coding for virus capsid protein. Virology, 167(1), 176-84 <br />
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The AAV capsid consists of 60 capsid protein subunits. The three cap proteins VP1, VP2, and VP3 are encoded in an overlapping reading frame. Arranged in a stoichiometric ratio of 1:1:10, they form an icosahedral symmetry. The mRNA encoding for the cap proteins is transcribed from p40 and alternative spliced to minor and major products. Alternative splicing and translation initiation of VP2 at a nonconventional ACG initiation codon promote the expression of VP1, VP2 and VP3. The VP proteins share a common C terminus and stop codon, but begin with a different start codon. The N termini of VP1 and VP2 play important roles in infection and contain motifs that are highly homologous to a phospholipase A2 (PLA2) domain and nuclear localization signals (NLSs).
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<center><img src="https://static.igem.org/mediawiki/parts/a/a7/Freiburg10_Cap_proteins_VP1_2%263.png" width="600"  
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height="auto"/></center>
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<b> Figure 1: The VP proteins are encoded in an overlapping open reading frame. </b>
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<center><img src="https://static.igem.org/mediawiki/parts/a/a7/Freiburg10_Cap_proteins_VP1_2%263.png" width="700"  
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height="auto" margin: 10px 10px 10px 10px/></center>
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<b> Figure 1: The VP proteins are encoded in an overlapping open reading frame. </b>.
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Revision as of 13:25, 27 October 2010

pCMV_[AAV2]-VP123


AAV2-VP123
Freiburg10 p5 promoter.png
BioBrick Nr. BBa_K404007
RFC standard RFC 25
Requirement pSB1C3
Source pAAV_RC from Stratagene
Submitted by [http://2010.igem.org/Team:Freiburg_Bioware FreiGEM 2010]

The AAV capsid consists of 60 capsid protein subunits. The three cap proteins VP1, VP2, and VP3 are encoded in an overlapping reading frame. Arranged in a stoichiometric ratio of 1:1:10, they form an icosahedral symmetry. The mRNA encoding for the cap proteins is transcribed from p40 and alternative spliced to minor and major products. Alternative splicing and translation initiation of VP2 at a nonconventional ACG initiation codon promote the expression of VP1, VP2 and VP3. The VP proteins share a common C terminus and stop codon, but begin with a different start codon. The N termini of VP1 and VP2 play important roles in infection and contain motifs that are highly homologous to a phospholipase A2 (PLA2) domain and nuclear localization signals (BR)(+). (Quelle)

Characterization

References

Cassinotti, P., Weitzand, M., & Tratschin, J. D., 1988. Organization of the adeno-associated virus (AAV) capsid gene: mapping of a minor spliced mRNA coding for virus capsid protein. Virology, 167(1), 176-84
Figure 1: The VP proteins are encoded in an overlapping open reading frame.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1729
    Illegal XhoI site found at 31
    Illegal XhoI site found at 217
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 1166