Difference between revisions of "Part:BBa K404241"

Revision as of 23:38, 27 October 2010 (view source) Kira M (Talk | contribs) ← Older edit		Revision as of 03:03, 28 October 2010 (view source) Kerstin (Talk | contribs) Newer edit →
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	__NOTOC__		__NOTOC__
	<partinfo>BBa_K404241 short</partinfo>		<partinfo>BBa_K404241 short</partinfo>
		+	{| style="color:black" cellpadding="6" cellspacing="1" border="2" align="left"
		+	! colspan="2" style="background:#66bbff;"|[https://parts.igem.org/Part:BBa_K404241 pCMV_(AAV2)-VP123ex (ViralBrick-453-BAP)]
		+	|-
		+	|'''BioBrick Nr.'''
		+	|[https://parts.igem.org/Part:BBa_K404241 BBa_K404241]
		+	|-
		+	|'''RFC standard'''
		+	|[https://parts.igem.org/Help:Assembly_standard_25 RFC 25]
		+	|-
		+	|'''Requirement'''
		+	|pSB1C3<br>
		+	|-
		+	|'''Source'''
		+	|
		+	|-
		+	|'''Submitted by'''
		+	|[http://2010.igem.org/Team:Freiburg_Bioware FreiGEM 2010]
		+	|}
		+	<br/><br/><br/><br/><br/><br/><br/><br/><br/><br/><br/><br/><br/>
	<br><b>The Biotinylation Acceptor peptide motif, inserted into the 453 loop of [[Part:BBa_K404007|pCMV_[AAV2]-VP123]]</b>		<br><b>The Biotinylation Acceptor peptide motif, inserted into the 453 loop of [[Part:BBa_K404007|pCMV_[AAV2]-VP123]]</b>

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Revision as of 03:03, 28 October 2010

pCMV_[AAV2]VP123 (ViralBrick-453-BAP)

pCMV_(AAV2)-VP123ex (ViralBrick-453-BAP)
BioBrick Nr.	BBa_K404241
RFC standard	RFC 25
Requirement	pSB1C3
Source
Submitted by	[http://2010.igem.org/Team:Freiburg_Bioware FreiGEM 2010]

The Biotinylation Acceptor peptide motif, inserted into the 453 loop of pCMV_[AAV2]-VP123

Usage and Biology

Capsid

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 the phospholipase A2 (PLA2) domain and 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., June 2002. SPromoters 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