Difference between revisions of "Part:BBa K404111"
 
(4 intermediate revisions by the same user not shown)
Line 25: Line 25:
 
<b>Guanylate kinases  (GMKs)</b> are involved in the salvage pathway of <b>mono-phosphorylated guanosine (GMP) </b>nucleosides to GDP therefore being essential in nucleotide maturation (Stolworthy & Black 2001). By introducing transgenic thymidine kinases (TKs) into tumor cells, a bottleneck occurs by overexpression of mono-phosphorylated intermediates. To overcome the accumulation of these non-toxic molecules, Willmon, Krabbenhoft, & Black (2006) fused the herpes simplex virus thymidine kinase (HSV-TK) to the guanylate kinase from M. musculus and demonstrated enhanced tumor killing in vitro. <br />
 
<b>Guanylate kinases  (GMKs)</b> are involved in the salvage pathway of <b>mono-phosphorylated guanosine (GMP) </b>nucleosides to GDP therefore being essential in nucleotide maturation (Stolworthy & Black 2001). By introducing transgenic thymidine kinases (TKs) into tumor cells, a bottleneck occurs by overexpression of mono-phosphorylated intermediates. To overcome the accumulation of these non-toxic molecules, Willmon, Krabbenhoft, & Black (2006) fused the herpes simplex virus thymidine kinase (HSV-TK) to the guanylate kinase from M. musculus and demonstrated enhanced tumor killing in vitro. <br />
 
<br />
 
<br />
−
https://static.igem.org/mediawiki/parts/a/a8/Freiburg10_GuanylateKinase.png <b>Figure 1: </b>Crystal structure (Sekulic et al. 2002)  of mouse guanylate kinase in complex with GMP and ADP (PDB: 1LVG).
+
<html>
 +
<img src="https://static.igem.org/mediawiki/parts/a/a8/Freiburg10_GuanylateKinase.png" style="margin: 0px 30px 0px 0px;" />
 +
 
 +
<b> Figure 1: </b>Crystal structure (Sekulic et al. 2002)  of mouse guanylate kinase in complex with GMP and ADP (PDB: 1LVG).
 
<br />
 
<br />
 
<br />
 
<br />
 
<br />
 
<br />
 
The <b>iGEM team Freiburg_Bioware</b> provides the mGMK BioBrick part within the <b>‘Virus Construction Kit’</b> for therapeutical applications. Producing viral particles with encapsidated single-stranded DNA containing the fusion gene mGMK_TK enhances the efficacy of the prodrug-activating system leading to cell death of targeted tumor cells
 
The <b>iGEM team Freiburg_Bioware</b> provides the mGMK BioBrick part within the <b>‘Virus Construction Kit’</b> for therapeutical applications. Producing viral particles with encapsidated single-stranded DNA containing the fusion gene mGMK_TK enhances the efficacy of the prodrug-activating system leading to cell death of targeted tumor cells
 +
</p>
 
<br />
 
<br />
 +
</div></html>
 
[[Image:Freiburg10_VectorplasmidBricks 8.png|thumb|center|480px]]<br>
 
[[Image:Freiburg10_VectorplasmidBricks 8.png|thumb|center|480px]]<br>
−
</p>
+
 
  
  
Line 47: Line 52:
 
<partinfo>BBa_K404111 parameters</partinfo>
 
<partinfo>BBa_K404111 parameters</partinfo>
 
<!-- -->
 
<!-- -->
 +
 +
<html>
 +
<h3>References</h3>
 +
<b>Sekulic, N. et al.</b>, 2002. Structural characterization of the closed conformation of mouse guanylate kinase. The Journal of biological chemistry, 277(33), 30236-43. Available at: http://www.ncbi.nlm.nih.gov/pubmed/12036965.
 +
<br />
 +
<b>Stolworthy, T.S. & Black, M.E.</b>, 2001. The mouse guanylate kinase double mutant E72Q/D103N is a functional adenylate kinase. Protein engineering, 14(11), 903-9. Available at: http://www.ncbi.nlm.nih.gov/pubmed/11742110.
 +
<br />
 +
<b>Willmon, C.L., Krabbenhoft, E. & Black, M.E.</b>, 2006. A guanylate kinase/HSV-1 thymidine kinase fusion protein enhances prodrug-mediated cell killing. Gene therapy, 13(17), 1309-12. Available at: http://www.ncbi.nlm.nih.gov/pubmed/16810197.
 +
</html>

Latest revision as of 22:07, 22 October 2010

Mouse guanylate kinase (mGMK)

Guanylate kinase (mGMK)
Freiburg10 VectorplasmidBricks 8.png
BioBrick Nr. K404111 BBa K404111
RFC standard RFC 25
Requirement pSB1C3
Source synthetic
Submitted by [http://2010.igem.org/Team:Freiburg_Bioware FreiGEM 2010]

Guanylate kinases (GMKs) are involved in the salvage pathway of mono-phosphorylated guanosine (GMP) nucleosides to GDP therefore being essential in nucleotide maturation (Stolworthy & Black 2001). By introducing transgenic thymidine kinases (TKs) into tumor cells, a bottleneck occurs by overexpression of mono-phosphorylated intermediates. To overcome the accumulation of these non-toxic molecules, Willmon, Krabbenhoft, & Black (2006) fused the herpes simplex virus thymidine kinase (HSV-TK) to the guanylate kinase from M. musculus and demonstrated enhanced tumor killing in vitro.

Figure 1: Crystal structure (Sekulic et al. 2002) of mouse guanylate kinase in complex with GMP and ADP (PDB: 1LVG).


The iGEM team Freiburg_Bioware provides the mGMK BioBrick part within the ‘Virus Construction Kit’ for therapeutical applications. Producing viral particles with encapsidated single-stranded DNA containing the fusion gene mGMK_TK enhances the efficacy of the prodrug-activating system leading to cell death of targeted tumor cells


Freiburg10 VectorplasmidBricks 8.png


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 SapI.rc site found at 45


References

Sekulic, N. et al., 2002. Structural characterization of the closed conformation of mouse guanylate kinase. The Journal of biological chemistry, 277(33), 30236-43. Available at: http://www.ncbi.nlm.nih.gov/pubmed/12036965.
Stolworthy, T.S. & Black, M.E., 2001. The mouse guanylate kinase double mutant E72Q/D103N is a functional adenylate kinase. Protein engineering, 14(11), 903-9. Available at: http://www.ncbi.nlm.nih.gov/pubmed/11742110.
Willmon, C.L., Krabbenhoft, E. & Black, M.E., 2006. A guanylate kinase/HSV-1 thymidine kinase fusion protein enhances prodrug-mediated cell killing. Gene therapy, 13(17), 1309-12. Available at: http://www.ncbi.nlm.nih.gov/pubmed/16810197.