Difference between revisions of "Part:BBa K404256"

Revision as of 16:58, 31 October 2010 (view source) Asi mit Niwoh (Talk | contribs) ← Older edit		Latest revision as of 16:38, 13 January 2011 (view source) Hanna (Talk | contribs)
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	|'''RFC standard'''		|'''RFC standard'''
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	|'''Requirement'''		|'''Requirement'''

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Latest revision as of 16:38, 13 January 2011

pCMV_[AAV2]-VP123ex (ViralBrick-587KO-Z34C-Spacer)
The Z34C antibody binding motif including a knockout of the natural HSPG tropism & spacer BBa_K404214 , inserted into the 453 loop of pCMV_[AAV2]-VP123

[pCMV_[AAV2-VP123ex(ViralBrick-587KO-Z34C-Spacer]
BioBrick Nr.	BBa_K404256
RFC standard	RFC 10
Requirement	pSB1C3
Source	pAAV_MCS provided by Stratagene
Submitted by	[http://2010.igem.org/Team:Freiburg_Bioware FreiGEM 2010]

Usage and Biology

[AAV2]-VP123ex

The pSB1C3_001_CMV_VP123 contains a CMV promoter upstream the VP123 sequence. 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 terminus of VP1 contains a motif that is highly homologous to the phospholipase A2 (PLA2) domain, both VP1 and VP2 have nuclear localization signals (BR)(+).
CMV promoter is derived from human Cytomegalovirus, which belongs to Herpesvirus group. All family members share the ability to remain in latent stage in the human body. CMV is located upstream of immediate-early gene. However, CMV promoter is an example of widely used promoters and is present in mammalian expression vectors. The advantage of CMV is the high-level constitutive expression in mostly all human tissues [Fitzsimons et al., 2002].

References

DiPrimio, Asokan, Govindasamy, Agbandje-McKenna, & Samulski, June 2008. Surface loop dynamics in adeno-associated virus capsid assembly. Journal of virology, 167(1), 5178–5189
Fitzsimons, H.L., Bland, R.J. & During, M.J. 2002. Promoters and regulatory elements that improve adeno-associated virus transgene expression in the brain. Methods San Diego Calif, 28(2), pp.227-236. Available at: http://www.ncbi.nlm.nih.gov/pubmed/12413421.
Figure 1: The VP proteins are encoded in an overlapping open reading frame.

Sequence and Features