Project

Part:BBa_K404210

Designed by: Freiburg Bioware 2010   Group: iGEM10_Freiburg_Bioware   (2010-08-28)
Revision as of 18:55, 26 October 2010 by Achim (Talk | contribs) (→‎Usage and Biology)

ViralBrick-587KO-Empty

Freiburg10 ViralBrick-logo-587KO-empty.png

Usage and Biology

The primary receptor of AAV-2 is the heparan sulfate proteoglycan (HSPG) receptor (Perabo et al. 2006). Its binding motif consists of five amino-acids located on the capsid surface: R484/R487, K532, R585/587. (Trepel et al. 2009). The positively charged arginine residues interact with the HSPGs' negatively charged acid residues. Opie et al. have shown that two point mutations (R585A and R588A) are sufficient to eliminate the heparin binding affinity in AAV2. (Opie et al. 2003). This ViralBrick has been created to introduce this knockout into other constructs. The biobricks with containing this knockout are annotated with „HSPG-ko“.


Restriction sites

Restriction site Enzyme Recognition sequence Activity Temp.
upstream 587 [http://www.neb.com/nebecomm/products/productR3136.asp BamHI-HF] Freiburg10 recognition site BamHI-HF.Gif 100;50;10;100 37°C
downstream 587 [http://www.neb.com/nebecomm/products/productR3151.asp PvuII-HF] Freiburg10 recognition site PvuII-HF.Gif 0;25;0;100 37°C

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 10
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
//chassis/eukaryote/human
//viral_vectors
//viral_vectors/aav
//viral_vectors/aav/capsid_coding
Parameters
None