Difference between revisions of "Part:BBa K1604030"
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<div style="text-align:center"><html><img src="https://static.igem.org/mediawiki/parts/e/ed/Unitn_pics_2015_NADpathway.png" style="width:75%;"></img></div></html> | <div style="text-align:center"><html><img src="https://static.igem.org/mediawiki/parts/e/ed/Unitn_pics_2015_NADpathway.png" style="width:75%;"></img></div></html> | ||
<p style="width:600px; margin-left:150px; margin-bottom:60px; | <p style="width:600px; margin-left:150px; margin-bottom:60px; | ||
− | text-align:justify "><b>FIGURE 1. Biochemical pathway of NAD synthesis.</b> PncB gene encodes the transcription of the NARPTase which catalyzes the formation of nicotinate mono-nucleotide from nicotinic acid. This is considered the limiting step of the synthesis of NAD. </p> | + | text-align:justify "><b>FIGURE 1. Biochemical pathway of NAD synthesis.</b> PncB gene encodes the transcription of the NARPTase which catalyzes the formation of nicotinate mono-nucleotide from nicotinic acid. This is considered the limiting step of the synthesis of NAD+. </p> |
− | <div style="text-align:center"><html><img src="https://static.igem.org/mediawiki/parts/b/b3/Unitn_pics_2015_WellRatioNAD | + | <div style="text-align:center"><html><img src="https://static.igem.org/mediawiki/parts/b/b3/Unitn_pics_2015_WellRatioNAD/NADH.png" style="width:85%;"></img></div></html> |
<p style="width:600px; margin-left:150px; margin-bottom:60px; | <p style="width:600px; margin-left:150px; margin-bottom:60px; | ||
− | text-align:justify "><b>FIGURE 2. PncB increases NAD levels by 126% (2.5 fold) and NADH levels by 44% (1.4 fold) when expressed in NEB10β.</b> NAD and NADH levels were calculated with a colorimetric assay using the Sigma NAD /NADH quantification kit (<html><a href="http://www.sigmaaldrich.com/catalog/product/sigma/mak037?lang=it®ion=IT">MAK037</a></html>) following the instructions described in the technical bulletin. The kit provides the measures of NAD levels indirectly from total levels of NAD + NADH and NADH only. Panel A. Colorimetric assay for NAD/NADH quantification. Panel B. NAD/NADH levels for three biological samples expressing <html><a href="https://parts.igem.org/Part:BBa_K1604031">BBa_K1604031</a></html> and one negative control expressing <html><a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K731201">BBa_K731201</a></html>. </p> | + | text-align:justify "><b>FIGURE 2. PncB increases NAD+ levels by 126% (2.5 fold) and NADH levels by 44% (1.4 fold) when expressed in NEB10β.</b> NAD+ and NADH levels were calculated with a colorimetric assay using the Sigma NAD/NADH quantification kit (<html><a href="http://www.sigmaaldrich.com/catalog/product/sigma/mak037?lang=it®ion=IT">MAK037</a></html>) following the instructions described in the technical bulletin. The kit provides the measures of NAD+ levels indirectly from total levels of NAD + NADH and NADH only. Panel A. Colorimetric assay for NAD/NADH quantification. Panel B. NAD/NADH levels for three biological samples expressing <html><a href="https://parts.igem.org/Part:BBa_K1604031">BBa_K1604031</a></html> and one negative control expressing <html><a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K731201">BBa_K731201</a></html>. </p> |
<b>For more information about the pncB (BBa_K1604030) characterization go on <html><a href="https://parts.igem.org/Part:BBa_K1604031">BBa_K1604031</a></html> page, or on Unitn iGEM 2015 project checking out our Wiki: <html><a href="http://2015.igem.org/Team:UNITN-Trento">Unitn iGEM wiki 2015.</a></p><html></b> | <b>For more information about the pncB (BBa_K1604030) characterization go on <html><a href="https://parts.igem.org/Part:BBa_K1604031">BBa_K1604031</a></html> page, or on Unitn iGEM 2015 project checking out our Wiki: <html><a href="http://2015.igem.org/Team:UNITN-Trento">Unitn iGEM wiki 2015.</a></p><html></b> |
Revision as of 10:04, 11 September 2015
pncB
pncB encodes for the enzyme NAPRTase (nicotinic acid phosphoribosyl-transferase). It catalyzes the formation of nicotinate mono-nucleotide, a direct precursor of NAD, from NA.[1] [2] [3].
Usage and Biology
FIGURE 1. Biochemical pathway of NAD synthesis. PncB gene encodes the transcription of the NARPTase which catalyzes the formation of nicotinate mono-nucleotide from nicotinic acid. This is considered the limiting step of the synthesis of NAD+.
FIGURE 2. PncB increases NAD+ levels by 126% (2.5 fold) and NADH levels by 44% (1.4 fold) when expressed in NEB10β. NAD+ and NADH levels were calculated with a colorimetric assay using the Sigma NAD/NADH quantification kit (MAK037) following the instructions described in the technical bulletin. The kit provides the measures of NAD+ levels indirectly from total levels of NAD + NADH and NADH only. Panel A. Colorimetric assay for NAD/NADH quantification. Panel B. NAD/NADH levels for three biological samples expressing BBa_K1604031 and one negative control expressing BBa_K731201.
For more information about the pncB (BBa_K1604030) characterization go on BBa_K1604031 page, or on Unitn iGEM 2015 project checking out our Wiki: Unitn iGEM wiki 2015.
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Susana J. Berrıos-Rivera, K.-Y. San, and G. N. Bennett. (2002) Metabolic Engineering 4, 238–247
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Marcel G. Wubbolts, Peter Terpstra, Jan B. van Beilen, Jaap Kingma, H. A. Rene Meesters, and Bernard Witholt. (1990) JBC, 265 (29), 17665-72
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Ka-Yiu San, George N. Bennett, Susana J. Berríos-Rivera, Ravi V. Vadali, Yea-Tyng Yang, Emily Horton, Fred B. Rudolph, Berna Sariyar, and Kimathi Blackwood (2002) Metabolic Engineering, 4, 182–192
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 1106
- 1000COMPATIBLE WITH RFC[1000]