Regulatory

Part:BBa_K404103

Designed by: Freiburg Bioware 2010   Group: iGEM10_Freiburg_Bioware   (2010-08-28)
Revision as of 02:51, 25 October 2010 by Bea (Talk | contribs)

p5 promoter

p5 promoter
Freiburg10 p5 promoter.png
BioBrick Nr. BBa_K404103
RFC standard RFC 10
Requirement pSB1C3
Source pAAV_RC from Stratagene
Submitted by [http://2010.igem.org/Team:Freiburg_Bioware FreiGEM 2010]

The p5 promoter, located downstream of the rep and cap ORF (Figure 5: The p5 promoter of the wtAAV-2 is located upstream of the rep and cap ORF and contains several elements, which interact with Rep and endogenous proteins.)of the wtAAV-2, regulates gene expression of the two larger non-structural proteins Rep 78 and Rep 68 that are essential in genome replication and viral genome integration into several hotspots of the human chromosome.
Several binding elements for cellular and viral proteins involved in regulation can be found in the p5 promoter(Figure 1) therefore playing an important role in gene transcription, integration and replication, dependent on the presence or absence of helper viruses such as adenovirus (Ad) or herpes simplex virus (HSV) (Murphy et al. 2007). Besides regulation of gene expression, the p5 integration efficient element (p5IEE) containing the rep binding element (RBE) and a terminal resolution site (trs) is responsible for mediating site specific integration into the human genome (Philpott et al. 2002).

Figure 1: The p5 promoter of the wtAAV-2 is located upstream of the rep and cap ORF and contains
several elements, which interact with Rep and endogenous proteins
.


Containing two consensus sequences for binding immediate early E1A gene product from adenoviruses (Chang et al. 1989), p5 promoter is transactivated in the presence of helper viruses whereas suppression occurs in absence of adenoviral proteins by low levels of Rep proteins (Beaton et al. 1989). Regulating of Rep78/68 by its negative feedback loop is critical since overexpression leads to cell cycle arrest in the S-phase (Berthet et al. 2005) and suppression of cellular promoters (Jing et al. 2001).


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
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 136


[edit]
Categories
//chassis/eukaryote/human
//function/regulation/transcriptional
//promoter
//viral_vectors
//viral_vectors/aav
//viral_vectors/aav/miscellaneous
Parameters
None