Part:BBa_K404105
p40 promoter
| p40 promoter | |
|---|---|
| BioBrick Nr. | BBa_K404105 |
| RFC standard | RFC 10 |
| Requirement | pSB1C3 |
| Source | |
| Submitted by | [http://2010.igem.org/Team:Freiburg_Bioware FreiGEM 2010] |
The P40 promoter is derived from the adeno-associated virus serotype 2 (AAV2) genome, where it is located at 40 map units. It regulates the transcription of the capsid proteins VP1, VP2 and VP3(Labow, Hermonat, & Berns, 1986; Cassinotti, Weitzand, & Tratschin, 1988). Several sequence regions have been identified to be important for maximal promoter activity: Two Sp1 sites, which are located 250 (Sp1-50) and 270 (GGT-70) base pairs upstream of the transcriptional start point and to which Sp1 or Sp1-like proteins bind (Pereira & Muzyczka 1997). Referring to the virus genome, p40 can also be induced through transactivation by the Rep proteins. The Sp1-50, together with the CArG-140 site of the P19 promoter, are the main elements involved in this process. The Rep proteins, which bind to the Rep binding element in the terminal repeat or the P5 promoter, can induce P19 or P40 by interaction with their bound Sp1 proteins thereby forming a DNA-loop (Pereira & Muzyczka 1997). In addition to that, the TATA box, located at 230, is also required for P40 activity. Furthermore the ATF-80 and the AP1-40 elements are also important for maximal promoter induction (Pereira & Muzyczka 1997).
Characterization
The Freiburg iGEM team 2010 characterized the P40 BioBrick by determining is promoter strength in comparison to the AAV2 p5 and p5_TATAless and the CMV promoter. This allows exact titration of plasmids for transfection and production of recombinant AAV2 particles.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
References
Cassinotti, P., Weitzand, M., & Tratschin, J. D., 1988. Organization of the adeno-associated virus (AAV) capsid gene: mapping of a minor spliced mRNA coding for virus capsid protein. Virology, 167(1), 176-84
Labow, M. A., Hermonat, P. L., & Berns, K. I., 1986. Positive and negative autoregulation of the adeno-associated virus type 2 genome. Journal of virology, 60(1), 251-8.
Pereira, D. J., & Muzyczka, N. , 1997. IThe adeno-associated virus type 2 p40 promoter requires a proximal Sp1 interaction and a p19 CArG-like element to facilitate Rep transactivation. Journal of virology, 71(6), 4300-9
//function/regulation/transcriptional
//promoter
//viral_vectors
//viral_vectors/aav
//viral_vectors/aav/miscellaneous
| None |