Difference between revisions of "Part:BBa K404011"
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__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K404011 short</partinfo>
 
<partinfo>BBa_K404011 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_K404011 pCMV_AAV2-VP2]
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|-
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|'''BioBrick Nr.'''
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|[https://parts.igem.org/Part:BBa_K404011 BBa_K404011]
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|-
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|'''RFC standard'''
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|[https://parts.igem.org/Help:Assembly_standard_10 RFC 10]
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|-
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|'''Requirement'''
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|pSB1C3_001<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 viral capsid is formed by the three structural proteins VP1, VP2 and VP3 which are encoded by the cap gene in an overlapping reading frame. They form an icosahedral symmetry arranged in a stoichiometric ratio of 1:1:10. The mRNA encoding for the cap proteins is transcribed from p40 and alternative spliced to minor and major products. Sharing a common C terminus and stop codon, the VP proteins begin with a different start codon. The translation of VP2 from the major spliced mRNA is not as efficient as of VP1 and VP3 because it initiates at a Thr codon (ACG). The N terminus of VP2 contains a motiv of about 70 amino acids that is highly homologous to a phospholipase A2 (PLA2) domain.  Furthermore, there are nuclear localization sequences (BR)(+) which are supposed to be necessary for endosomal escape and nuclear entry. The exact role of VP2 remains unknown, although the protein is thought to be nonessential for viral assembly and infectivity.
  
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<h1>Usage and Biology</h1>
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(Quelle)
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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 translation of VP2 from the major spliced mRNA is not as efficient as of VP1 and VP3 because it initiates at a Thr codon (ACG). The N terminus of VP1 has an extension of 65 amino acids and similar to VP1, it contains two functional elements: a phospholipase A2 (PLA2) domain and nuclear localization signals (BR)(+). These elements are conserved almost in all parvoviruses. The exact role of VP2 remains unknown, although the protein is thought to be nonessential for viral assembly and infectivity.
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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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<h3>Characterization</h3>
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<h3>References</h3>
  
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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" margin: 10px 10px 10px 10px/></center>
 
<b> Figure 1: The VP proteins are encoded in an overlapping open reading frame. </b>.
 
<b> Figure 1: The VP proteins are encoded in an overlapping open reading frame. </b>.
 
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>

Revision as of 13:50, 27 October 2010

pCMV_[AAV2]-VP2

pCMV_AAV2-VP2
BioBrick Nr. BBa_K404011
RFC standard RFC 10
Requirement pSB1C3_001
Source pAAV_RC from Stratagene
Submitted by [http://2010.igem.org/Team:Freiburg_Bioware FreiGEM 2010]

The viral capsid is formed by the three structural proteins VP1, VP2 and VP3 which are encoded by the cap gene in an overlapping reading frame. They form an icosahedral symmetry arranged in a stoichiometric ratio of 1:1:10. The mRNA encoding for the cap proteins is transcribed from p40 and alternative spliced to minor and major products. Sharing a common C terminus and stop codon, the VP proteins begin with a different start codon. The translation of VP2 from the major spliced mRNA is not as efficient as of VP1 and VP3 because it initiates at a Thr codon (ACG). The N terminus of VP2 contains a motiv of about 70 amino acids that is highly homologous to a phospholipase A2 (PLA2) domain. Furthermore, there are nuclear localization sequences (BR)(+) which are supposed to be necessary for endosomal escape and nuclear entry. The exact role of VP2 remains unknown, although the protein is thought to be nonessential for viral assembly and infectivity. (Quelle)

Characterization

References

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
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 665
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 1425