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Team:Alberta-North-RBI E/genetics
From 2012e.igem.org
(Difference between revisions)
(→Genetics) |
(→Genetics) |
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<center> | <center> | ||
BioBricks to be made: | BioBricks to be made: | ||
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| - | + | ||
| - | + | ||
<table class=MsoNormalTable border=0 cellspacing=0 cellpadding=0 | <table class=MsoNormalTable border=0 cellspacing=0 cellpadding=0 | ||
| Line 618: | Line 616: | ||
</table> | </table> | ||
| + | <center> | ||
| + | Genes to be knocked out: | ||
| + | |||
| + | |||
| + | <table class=MsoNormalTable border=0 cellspacing=0 cellpadding=0 | ||
| + | style='border-collapse:collapse;mso-padding-alt:0cm 5.4pt 0cm 5.4pt'> | ||
| + | <tr style='height:15.75pt'> | ||
| + | <td width=216 valign=bottom style='width:162.0pt;border-top:solid black 1.0pt; | ||
| + | border-left:solid black 1.0pt;border-bottom:none;border-right:none; | ||
| + | mso-border-top-alt:solid black .5pt;mso-border-left-alt:solid black 1.0pt; | ||
| + | background:white;padding:0cm 5.4pt 0cm 5.4pt;height:15.75pt'> | ||
| + | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;mso-line-height-alt: | ||
| + | 5.0pt'><span class=SpellE><span lang=EN-US style='font-size:10.0pt; | ||
| + | mso-fareast-font-family:"Times New Roman";color:black'>Shikimate</span></span><span | ||
| + | lang=EN-US style='font-size:10.0pt;mso-fareast-font-family:"Times New Roman"; | ||
| + | color:black'> kinase<o:p></o:p></span></p> | ||
| + | </td> | ||
| + | <td width=68 valign=bottom style='width:50.95pt;border:none;border-top:solid black 1.0pt; | ||
| + | mso-border-top-alt:solid black .5pt;background:white;padding:0cm 5.4pt 0cm 5.4pt; | ||
| + | height:15.75pt'> | ||
| + | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;mso-line-height-alt: | ||
| + | 5.0pt'><span lang=EN-US style='font-size:10.0pt;mso-fareast-font-family:"Times New Roman"; | ||
| + | color:black'>PPS_4924<i><o:p></o:p></i></span></p> | ||
| + | </td> | ||
| + | <td width=74 valign=bottom style='width:55.25pt;border:none;border-top:solid black 1.0pt; | ||
| + | mso-border-top-alt:solid black .5pt;background:white;padding:0cm 5.4pt 0cm 5.4pt; | ||
| + | height:15.75pt'> | ||
| + | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;mso-line-height-alt: | ||
| + | 5.0pt'><i><span lang=EN-US style='font-size:10.0pt;mso-fareast-font-family: | ||
| + | "Times New Roman";color:black'>P. <span class=SpellE>putida</span></span></i><span | ||
| + | lang=EN-US style='font-size:10.0pt;mso-fareast-font-family:"Times New Roman"; | ||
| + | color:black'><o:p></o:p></span></p> | ||
| + | </td> | ||
| + | <td width=275 valign=bottom style='width:205.9pt;border-top:solid black 1.0pt; | ||
| + | border-left:none;border-bottom:none;border-right:solid black 1.0pt; | ||
| + | mso-border-top-alt:solid black .5pt;mso-border-right-alt:solid black 1.0pt; | ||
| + | background:white;padding:0cm 5.4pt 0cm 5.4pt;height:15.75pt'> | ||
| + | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;mso-line-height-alt: | ||
| + | 5.0pt'><span lang=EN-US style='font-size:10.0pt;mso-fareast-font-family:"Times New Roman"; | ||
| + | color:black'>Removed and reintroduced as inducible<o:p></o:p></span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr style='height:15.75pt'> | ||
| + | <td width=216 valign=bottom style='width:162.0pt;border-top:solid black 1.0pt; | ||
| + | border-left:solid black 1.0pt;border-bottom:none;border-right:none; | ||
| + | mso-border-top-alt:solid black .5pt;mso-border-left-alt:solid black 1.0pt; | ||
| + | background:white;padding:0cm 5.4pt 0cm 5.4pt;height:15.75pt'> | ||
| + | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;mso-line-height-alt: | ||
| + | 5.0pt'><span lang=EN-US style='font-size:10.0pt;mso-fareast-font-family:"Times New Roman"; | ||
| + | color:black'>3-phosphoshikimate 1-carboxyvinyltransferase<o:p></o:p></span></p> | ||
| + | </td> | ||
| + | <td width=68 valign=bottom style='width:50.95pt;border:none;border-top:solid black 1.0pt; | ||
| + | mso-border-top-alt:solid black .5pt;background:white;padding:0cm 5.4pt 0cm 5.4pt; | ||
| + | height:15.75pt'> | ||
| + | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;mso-line-height-alt: | ||
| + | 5.0pt'><span lang=EN-US style='font-size:10.0pt;mso-fareast-font-family:"Times New Roman"; | ||
| + | color:black'>PPS_1411<i><o:p></o:p></i></span></p> | ||
| + | </td> | ||
| + | <td width=74 valign=bottom style='width:55.25pt;border:none;border-top:solid black 1.0pt; | ||
| + | mso-border-top-alt:solid black .5pt;background:white;padding:0cm 5.4pt 0cm 5.4pt; | ||
| + | height:15.75pt'> | ||
| + | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;mso-line-height-alt: | ||
| + | 5.0pt'><i><span lang=EN-US style='font-size:10.0pt;mso-fareast-font-family: | ||
| + | "Times New Roman";color:black'>P. <span class=SpellE>putida</span></span></i><span | ||
| + | lang=EN-US style='font-size:10.0pt;mso-fareast-font-family:"Times New Roman"; | ||
| + | color:black'><o:p></o:p></span></p> | ||
| + | </td> | ||
| + | <td width=275 valign=bottom style='width:205.9pt;border-top:solid black 1.0pt; | ||
| + | border-left:none;border-bottom:none;border-right:solid black 1.0pt; | ||
| + | mso-border-top-alt:solid black .5pt;mso-border-right-alt:solid black 1.0pt; | ||
| + | background:white;padding:0cm 5.4pt 0cm 5.4pt;height:15.75pt'> | ||
| + | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;mso-line-height-alt: | ||
| + | 5.0pt'><span lang=EN-US style='font-size:10.0pt;mso-fareast-font-family:"Times New Roman"; | ||
| + | color:black'>Removed and reintroduced as inducible<o:p></o:p></span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr style='height:15.75pt'> | ||
| + | <td width=216 valign=bottom style='width:162.0pt;border-top:solid black 1.0pt; | ||
| + | border-left:solid black 1.0pt;border-bottom:none;border-right:none; | ||
| + | mso-border-top-alt:solid black .5pt;mso-border-left-alt:solid black 1.0pt; | ||
| + | background:white;padding:0cm 5.4pt 0cm 5.4pt;height:15.75pt'> | ||
| + | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;mso-line-height-alt: | ||
| + | 5.0pt'><span class=SpellE><span lang=EN-US style='font-size:10.0pt; | ||
| + | mso-fareast-font-family:"Times New Roman";color:black'>Chorismate</span></span><span | ||
| + | lang=EN-US style='font-size:10.0pt;mso-fareast-font-family:"Times New Roman"; | ||
| + | color:black'> synthase<o:p></o:p></span></p> | ||
| + | </td> | ||
| + | <td width=68 valign=bottom style='width:50.95pt;border:none;border-top:solid black 1.0pt; | ||
| + | mso-border-top-alt:solid black .5pt;background:white;padding:0cm 5.4pt 0cm 5.4pt; | ||
| + | height:15.75pt'> | ||
| + | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;mso-line-height-alt: | ||
| + | 5.0pt'><span lang=EN-US style='font-size:10.0pt;mso-fareast-font-family:"Times New Roman"; | ||
| + | color:black'>PPS_1471<i><o:p></o:p></i></span></p> | ||
| + | </td> | ||
| + | <td width=74 valign=bottom style='width:55.25pt;border:none;border-top:solid black 1.0pt; | ||
| + | mso-border-top-alt:solid black .5pt;background:white;padding:0cm 5.4pt 0cm 5.4pt; | ||
| + | height:15.75pt'> | ||
| + | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;mso-line-height-alt: | ||
| + | 5.0pt'><i><span lang=EN-US style='font-size:10.0pt;mso-fareast-font-family: | ||
| + | "Times New Roman";color:black'>P. <span class=SpellE>putida</span></span></i><span | ||
| + | lang=EN-US style='font-size:10.0pt;mso-fareast-font-family:"Times New Roman"; | ||
| + | color:black'><o:p></o:p></span></p> | ||
| + | </td> | ||
| + | <td width=275 valign=bottom style='width:205.9pt;border-top:solid black 1.0pt; | ||
| + | border-left:none;border-bottom:none;border-right:solid black 1.0pt; | ||
| + | mso-border-top-alt:solid black .5pt;mso-border-right-alt:solid black 1.0pt; | ||
| + | background:white;padding:0cm 5.4pt 0cm 5.4pt;height:15.75pt'> | ||
| + | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;mso-line-height-alt: | ||
| + | 5.0pt'><span lang=EN-US style='font-size:10.0pt;mso-fareast-font-family:"Times New Roman"; | ||
| + | color:black'>Removed and reintroduced as inducible<o:p></o:p></span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr style='height:15.75pt'> | ||
| + | <td width=216 valign=bottom style='width:162.0pt;border-top:solid black 1.0pt; | ||
| + | border-left:solid black 1.0pt;border-bottom:none;border-right:none; | ||
| + | mso-border-top-alt:solid black .5pt;mso-border-left-alt:solid black 1.0pt; | ||
| + | background:white;padding:0cm 5.4pt 0cm 5.4pt;height:15.75pt'> | ||
| + | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;mso-line-height-alt: | ||
| + | 5.0pt'><span class=SpellE><span lang=EN-US style='font-size:10.0pt; | ||
| + | mso-fareast-font-family:"Times New Roman";color:black'>Chorismatemutase</span></span><span | ||
| + | lang=EN-US style='font-size:10.0pt;mso-fareast-font-family:"Times New Roman"; | ||
| + | color:black'><o:p></o:p></span></p> | ||
| + | </td> | ||
| + | <td width=68 valign=bottom style='width:50.95pt;border:none;border-top:solid black 1.0pt; | ||
| + | mso-border-top-alt:solid black .5pt;background:white;padding:0cm 5.4pt 0cm 5.4pt; | ||
| + | height:15.75pt'> | ||
| + | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;mso-line-height-alt: | ||
| + | 5.0pt'><span lang=EN-US style='font-size:10.0pt;mso-fareast-font-family:"Times New Roman"; | ||
| + | color:black'>PPS_1410<i><o:p></o:p></i></span></p> | ||
| + | </td> | ||
| + | <td width=74 valign=bottom style='width:55.25pt;border:none;border-top:solid black 1.0pt; | ||
| + | mso-border-top-alt:solid black .5pt;background:white;padding:0cm 5.4pt 0cm 5.4pt; | ||
| + | height:15.75pt'> | ||
| + | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;mso-line-height-alt: | ||
| + | 5.0pt'><i><span lang=EN-US style='font-size:10.0pt;mso-fareast-font-family: | ||
| + | "Times New Roman";color:black'>P. <span class=SpellE>putida</span></span></i><span | ||
| + | lang=EN-US style='font-size:10.0pt;mso-fareast-font-family:"Times New Roman"; | ||
| + | color:black'><o:p></o:p></span></p> | ||
| + | </td> | ||
| + | <td width=275 valign=bottom style='width:205.9pt;border-top:solid black 1.0pt; | ||
| + | border-left:none;border-bottom:none;border-right:solid black 1.0pt; | ||
| + | mso-border-top-alt:solid black .5pt;mso-border-right-alt:solid black 1.0pt; | ||
| + | background:white;padding:0cm 5.4pt 0cm 5.4pt;height:15.75pt'> | ||
| + | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;mso-line-height-alt: | ||
| + | 5.0pt'><span lang=EN-US style='font-size:10.0pt;mso-fareast-font-family:"Times New Roman"; | ||
| + | color:black'>Removed and reintroduced as inducible<o:p></o:p></span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr style='height:15.75pt'> | ||
| + | <td width=216 valign=bottom style='width:162.0pt;border-top:solid black 1.0pt; | ||
| + | border-left:solid black 1.0pt;border-bottom:none;border-right:none; | ||
| + | mso-border-top-alt:solid black .5pt;mso-border-left-alt:solid black 1.0pt; | ||
| + | background:white;padding:0cm 5.4pt 0cm 5.4pt;height:15.75pt'> | ||
| + | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;mso-line-height-alt: | ||
| + | 5.0pt'><span lang=EN-US style='font-size:10.0pt;mso-fareast-font-family:"Times New Roman"; | ||
| + | color:black'>Aromatic amino acid aminotransferase<o:p></o:p></span></p> | ||
| + | </td> | ||
| + | <td width=68 valign=bottom style='width:50.95pt;border:none;border-top:solid black 1.0pt; | ||
| + | mso-border-top-alt:solid black .5pt;background:white;padding:0cm 5.4pt 0cm 5.4pt; | ||
| + | height:15.75pt'> | ||
| + | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;mso-line-height-alt: | ||
| + | 5.0pt'><span lang=EN-US style='font-size:10.0pt;mso-fareast-font-family:"Times New Roman"; | ||
| + | color:black'>PPS_1561<i><o:p></o:p></i></span></p> | ||
| + | </td> | ||
| + | <td width=74 valign=bottom style='width:55.25pt;border:none;border-top:solid black 1.0pt; | ||
| + | mso-border-top-alt:solid black .5pt;background:white;padding:0cm 5.4pt 0cm 5.4pt; | ||
| + | height:15.75pt'> | ||
| + | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;mso-line-height-alt: | ||
| + | 5.0pt'><i><span lang=EN-US style='font-size:10.0pt;mso-fareast-font-family: | ||
| + | "Times New Roman";color:black'>P. <span class=SpellE>putida</span></span></i><span | ||
| + | lang=EN-US style='font-size:10.0pt;mso-fareast-font-family:"Times New Roman"; | ||
| + | color:black'><o:p></o:p></span></p> | ||
| + | </td> | ||
| + | <td width=275 valign=bottom style='width:205.9pt;border-top:solid black 1.0pt; | ||
| + | border-left:none;border-bottom:none;border-right:solid black 1.0pt; | ||
| + | mso-border-top-alt:solid black .5pt;mso-border-right-alt:solid black 1.0pt; | ||
| + | background:white;padding:0cm 5.4pt 0cm 5.4pt;height:15.75pt'> | ||
| + | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;mso-line-height-alt: | ||
| + | 5.0pt'><span lang=EN-US style='font-size:10.0pt;mso-fareast-font-family:"Times New Roman"; | ||
| + | color:black'>Removed and reintroduced as inducible<o:p></o:p></span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr style='height:15.75pt'> | ||
| + | <td width=216 valign=bottom style='width:162.0pt;border-top:solid black 1.0pt; | ||
| + | border-left:solid black 1.0pt;border-bottom:none;border-right:none; | ||
| + | mso-border-top-alt:solid black .5pt;mso-border-left-alt:solid black 1.0pt; | ||
| + | background:white;padding:0cm 5.4pt 0cm 5.4pt;height:15.75pt'> | ||
| + | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;mso-line-height-alt: | ||
| + | 5.0pt'><span lang=EN-US style='font-size:10.0pt;mso-fareast-font-family:"Times New Roman"; | ||
| + | color:black'>Aromatic amino acid aminotransferase<o:p></o:p></span></p> | ||
| + | </td> | ||
| + | <td width=68 valign=bottom style='width:50.95pt;border:none;border-top:solid black 1.0pt; | ||
| + | mso-border-top-alt:solid black .5pt;background:white;padding:0cm 5.4pt 0cm 5.4pt; | ||
| + | height:15.75pt'> | ||
| + | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;mso-line-height-alt: | ||
| + | 5.0pt'><span lang=EN-US style='font-size:10.0pt;mso-fareast-font-family:"Times New Roman"; | ||
| + | color:black'>PPS_3083<i><o:p></o:p></i></span></p> | ||
| + | </td> | ||
| + | <td width=74 valign=bottom style='width:55.25pt;border:none;border-top:solid black 1.0pt; | ||
| + | mso-border-top-alt:solid black .5pt;background:white;padding:0cm 5.4pt 0cm 5.4pt; | ||
| + | height:15.75pt'> | ||
| + | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;mso-line-height-alt: | ||
| + | 5.0pt'><i><span lang=EN-US style='font-size:10.0pt;mso-fareast-font-family: | ||
| + | "Times New Roman";color:black'>P. <span class=SpellE>putida</span></span></i><span | ||
| + | lang=EN-US style='font-size:10.0pt;mso-fareast-font-family:"Times New Roman"; | ||
| + | color:black'><o:p></o:p></span></p> | ||
| + | </td> | ||
| + | <td width=275 valign=bottom style='width:205.9pt;border-top:solid black 1.0pt; | ||
| + | border-left:none;border-bottom:none;border-right:solid black 1.0pt; | ||
| + | mso-border-top-alt:solid black .5pt;mso-border-right-alt:solid black 1.0pt; | ||
| + | background:white;padding:0cm 5.4pt 0cm 5.4pt;height:15.75pt'> | ||
| + | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;mso-line-height-alt: | ||
| + | 5.0pt'><span lang=EN-US style='font-size:10.0pt;mso-fareast-font-family:"Times New Roman"; | ||
| + | color:black'>Removed and reintroduced as inducible<o:p></o:p></span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr style='height:15.75pt'> | ||
| + | <td width=216 valign=bottom style='width:162.0pt;border-top:solid black 1.0pt; | ||
| + | border-left:solid black 1.0pt;border-bottom:none;border-right:none; | ||
| + | mso-border-top-alt:solid black .5pt;mso-border-left-alt:solid black 1.0pt; | ||
| + | background:white;padding:0cm 5.4pt 0cm 5.4pt;height:15.75pt'> | ||
| + | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;mso-line-height-alt: | ||
| + | 5.0pt'><span class=SpellE><span lang=EN-US style='font-size:10.0pt; | ||
| + | mso-fareast-font-family:"Times New Roman";color:black'>Histidinol</span></span><span | ||
| + | lang=EN-US style='font-size:10.0pt;mso-fareast-font-family:"Times New Roman"; | ||
| + | color:black'>-phosphate aminotransferase<o:p></o:p></span></p> | ||
| + | </td> | ||
| + | <td width=68 valign=bottom style='width:50.95pt;border:none;border-top:solid black 1.0pt; | ||
| + | mso-border-top-alt:solid black .5pt;background:white;padding:0cm 5.4pt 0cm 5.4pt; | ||
| + | height:15.75pt'> | ||
| + | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;mso-line-height-alt: | ||
| + | 5.0pt'><span lang=EN-US style='font-size:10.0pt;mso-fareast-font-family:"Times New Roman"; | ||
| + | color:black'>PPS_0995<i><o:p></o:p></i></span></p> | ||
| + | </td> | ||
| + | <td width=74 valign=bottom style='width:55.25pt;border:none;border-top:solid black 1.0pt; | ||
| + | mso-border-top-alt:solid black .5pt;background:white;padding:0cm 5.4pt 0cm 5.4pt; | ||
| + | height:15.75pt'> | ||
| + | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;mso-line-height-alt: | ||
| + | 5.0pt'><i><span lang=EN-US style='font-size:10.0pt;mso-fareast-font-family: | ||
| + | "Times New Roman";color:black'>P. <span class=SpellE>putida</span></span></i><span | ||
| + | lang=EN-US style='font-size:10.0pt;mso-fareast-font-family:"Times New Roman"; | ||
| + | color:black'><o:p></o:p></span></p> | ||
| + | </td> | ||
| + | <td width=275 valign=bottom style='width:205.9pt;border-top:solid black 1.0pt; | ||
| + | border-left:none;border-bottom:none;border-right:solid black 1.0pt; | ||
| + | mso-border-top-alt:solid black .5pt;mso-border-right-alt:solid black 1.0pt; | ||
| + | background:white;padding:0cm 5.4pt 0cm 5.4pt;height:15.75pt'> | ||
| + | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;mso-line-height-alt: | ||
| + | 5.0pt'><span lang=EN-US style='font-size:10.0pt;mso-fareast-font-family:"Times New Roman"; | ||
| + | color:black'>Removed and reintroduced as inducible<o:p></o:p></span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr style='height:15.75pt'> | ||
| + | <td width=216 valign=bottom style='width:162.0pt;border-top:solid black 1.0pt; | ||
| + | border-left:solid black 1.0pt;border-bottom:none;border-right:none; | ||
| + | mso-border-top-alt:solid black .5pt;mso-border-left-alt:solid black 1.0pt; | ||
| + | background:white;padding:0cm 5.4pt 0cm 5.4pt;height:15.75pt'> | ||
| + | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;mso-line-height-alt: | ||
| + | 5.0pt'><span class=SpellE><span lang=EN-US style='font-size:10.0pt; | ||
| + | mso-fareast-font-family:"Times New Roman";color:black'>Pyrroloquinolinequinone</span></span><span | ||
| + | lang=EN-US style='font-size:10.0pt;mso-fareast-font-family:"Times New Roman"; | ||
| + | color:black'><o:p></o:p></span></p> | ||
| + | </td> | ||
| + | <td width=68 valign=bottom style='width:50.95pt;border:none;border-top:solid black 1.0pt; | ||
| + | mso-border-top-alt:solid black .5pt;background:white;padding:0cm 5.4pt 0cm 5.4pt; | ||
| + | height:15.75pt'> | ||
| + | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;mso-line-height-alt: | ||
| + | 5.0pt'><span lang=EN-US style='font-size:10.0pt;mso-fareast-font-family:"Times New Roman"; | ||
| + | color:black'>PPS_3064<i><o:p></o:p></i></span></p> | ||
| + | </td> | ||
| + | <td width=74 valign=bottom style='width:55.25pt;border:none;border-top:solid black 1.0pt; | ||
| + | mso-border-top-alt:solid black .5pt;background:white;padding:0cm 5.4pt 0cm 5.4pt; | ||
| + | height:15.75pt'> | ||
| + | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;mso-line-height-alt: | ||
| + | 5.0pt'><i><span lang=EN-US style='font-size:10.0pt;mso-fareast-font-family: | ||
| + | "Times New Roman";color:black'>P. <span class=SpellE>putida</span></span></i><span | ||
| + | lang=EN-US style='font-size:10.0pt;mso-fareast-font-family:"Times New Roman"; | ||
| + | color:black'><o:p></o:p></span></p> | ||
| + | </td> | ||
| + | <td width=275 valign=bottom style='width:205.9pt;border-top:solid black 1.0pt; | ||
| + | border-left:none;border-bottom:none;border-right:solid black 1.0pt; | ||
| + | mso-border-top-alt:solid black .5pt;mso-border-right-alt:solid black 1.0pt; | ||
| + | background:white;padding:0cm 5.4pt 0cm 5.4pt;height:15.75pt'> | ||
| + | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;mso-line-height-alt: | ||
| + | 5.0pt'><span lang=EN-US style='font-size:10.0pt;mso-fareast-font-family:"Times New Roman"; | ||
| + | color:black'>Removed<o:p></o:p></span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr style='height:15.75pt'> | ||
| + | <td width=216 valign=bottom style='width:162.0pt;border:none;border-left: | ||
| + | solid black 1.0pt;background:white;padding:0cm 5.4pt 0cm 5.4pt;height:15.75pt'> | ||
| + | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;mso-line-height-alt: | ||
| + | 5.0pt'><span class=SpellE><span lang=EN-US style='font-size:10.0pt; | ||
| + | mso-fareast-font-family:"Times New Roman";color:black'>Anthranilate</span></span><span | ||
| + | lang=EN-US style='font-size:10.0pt;mso-fareast-font-family:"Times New Roman"; | ||
| + | color:black'> synthase component I<o:p></o:p></span></p> | ||
| + | </td> | ||
| + | <td width=68 valign=bottom style='width:50.95pt;background:white;padding: | ||
| + | 0cm 5.4pt 0cm 5.4pt;height:15.75pt'> | ||
| + | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;mso-line-height-alt: | ||
| + | 5.0pt'><span lang=EN-US style='font-size:10.0pt;mso-fareast-font-family:"Times New Roman"; | ||
| + | color:black'>PPS_0413<i><o:p></o:p></i></span></p> | ||
| + | </td> | ||
| + | <td width=74 valign=bottom style='width:55.25pt;background:white;padding: | ||
| + | 0cm 5.4pt 0cm 5.4pt;height:15.75pt'> | ||
| + | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;mso-line-height-alt: | ||
| + | 5.0pt'><i><span lang=EN-US style='font-size:10.0pt;mso-fareast-font-family: | ||
| + | "Times New Roman";color:black'>P. <span class=SpellE>putida</span></span></i><span | ||
| + | lang=EN-US style='font-size:10.0pt;mso-fareast-font-family:"Times New Roman"; | ||
| + | color:black'><o:p></o:p></span></p> | ||
| + | </td> | ||
| + | <td width=275 valign=bottom style='width:205.9pt;border:none;border-right: | ||
| + | solid black 1.0pt;background:white;padding:0cm 5.4pt 0cm 5.4pt;height:15.75pt'> | ||
| + | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;mso-line-height-alt: | ||
| + | 5.0pt'><span lang=EN-US style='font-size:10.0pt;mso-fareast-font-family:"Times New Roman"; | ||
| + | color:black'>Removed<o:p></o:p></span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr style='height:15.75pt'> | ||
| + | <td width=216 valign=bottom style='width:162.0pt;border:none;border-left: | ||
| + | solid black 1.0pt;background:white;padding:0cm 5.4pt 0cm 5.4pt;height:15.75pt'> | ||
| + | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;mso-line-height-alt: | ||
| + | 5.0pt'><span class=SpellE><span lang=EN-US style='font-size:10.0pt; | ||
| + | mso-fareast-font-family:"Times New Roman";color:black'>Anthranilate</span></span><span | ||
| + | lang=EN-US style='font-size:10.0pt;mso-fareast-font-family:"Times New Roman"; | ||
| + | color:black'> synthase component II<o:p></o:p></span></p> | ||
| + | </td> | ||
| + | <td width=68 valign=bottom style='width:50.95pt;background:white;padding: | ||
| + | 0cm 5.4pt 0cm 5.4pt;height:15.75pt'> | ||
| + | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;mso-line-height-alt: | ||
| + | 5.0pt'><span lang=EN-US style='font-size:10.0pt;mso-fareast-font-family:"Times New Roman"; | ||
| + | color:black'>PPS_0416<i><o:p></o:p></i></span></p> | ||
| + | </td> | ||
| + | <td width=74 valign=bottom style='width:55.25pt;background:white;padding: | ||
| + | 0cm 5.4pt 0cm 5.4pt;height:15.75pt'> | ||
| + | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;mso-line-height-alt: | ||
| + | 5.0pt'><i><span lang=EN-US style='font-size:10.0pt;mso-fareast-font-family: | ||
| + | "Times New Roman";color:black'>P. <span class=SpellE>putida</span></span></i><span | ||
| + | lang=EN-US style='font-size:10.0pt;mso-fareast-font-family:"Times New Roman"; | ||
| + | color:black'><o:p></o:p></span></p> | ||
| + | </td> | ||
| + | <td width=275 valign=bottom style='width:205.9pt;border:none;border-right: | ||
| + | solid black 1.0pt;background:white;padding:0cm 5.4pt 0cm 5.4pt;height:15.75pt'> | ||
| + | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;mso-line-height-alt: | ||
| + | 5.0pt'><span lang=EN-US style='font-size:10.0pt;mso-fareast-font-family:"Times New Roman"; | ||
| + | color:black'>Removed<o:p></o:p></span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr style='height:15.75pt'> | ||
| + | <td width=216 valign=bottom style='width:162.0pt;border:none;border-left: | ||
| + | solid black 1.0pt;background:white;padding:0cm 5.4pt 0cm 5.4pt;height:15.75pt'> | ||
| + | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;mso-line-height-alt: | ||
| + | 5.0pt'><span lang=EN-US style='font-size:10.0pt;mso-fareast-font-family:"Times New Roman"; | ||
| + | color:black'>Para-<span class=SpellE>aminobenzoate</span> synthase subunit I<o:p></o:p></span></p> | ||
| + | </td> | ||
| + | <td width=68 valign=bottom style='width:50.95pt;background:white;padding: | ||
| + | 0cm 5.4pt 0cm 5.4pt;height:15.75pt'> | ||
| + | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;mso-line-height-alt: | ||
| + | 5.0pt'><span lang=EN-US style='font-size:10.0pt;mso-fareast-font-family:"Times New Roman"; | ||
| + | color:black'>PPS_1911<i><o:p></o:p></i></span></p> | ||
| + | </td> | ||
| + | <td width=74 valign=bottom style='width:55.25pt;background:white;padding: | ||
| + | 0cm 5.4pt 0cm 5.4pt;height:15.75pt'> | ||
| + | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;mso-line-height-alt: | ||
| + | 5.0pt'><i><span lang=EN-US style='font-size:10.0pt;mso-fareast-font-family: | ||
| + | "Times New Roman";color:black'>P. <span class=SpellE>putida</span></span></i><span | ||
| + | lang=EN-US style='font-size:10.0pt;mso-fareast-font-family:"Times New Roman"; | ||
| + | color:black'><o:p></o:p></span></p> | ||
| + | </td> | ||
| + | <td width=275 valign=bottom style='width:205.9pt;border:none;border-right: | ||
| + | solid black 1.0pt;background:white;padding:0cm 5.4pt 0cm 5.4pt;height:15.75pt'> | ||
| + | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;mso-line-height-alt: | ||
| + | 5.0pt'><span lang=EN-US style='font-size:10.0pt;mso-fareast-font-family:"Times New Roman"; | ||
| + | color:black'>Removed<o:p></o:p></span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr style='height:15.75pt'> | ||
| + | <td width=216 valign=bottom style='width:162.0pt;border:none;border-left: | ||
| + | solid black 1.0pt;background:white;padding:0cm 5.4pt 0cm 5.4pt;height:15.75pt'> | ||
| + | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;mso-line-height-alt: | ||
| + | 5.0pt'><span class=SpellE><span lang=EN-US style='font-size:10.0pt; | ||
| + | mso-fareast-font-family:"Times New Roman";color:black'>Shikimate</span></span><span | ||
| + | lang=EN-US style='font-size:10.0pt;mso-fareast-font-family:"Times New Roman"; | ||
| + | color:black'> transporter<o:p></o:p></span></p> | ||
| + | </td> | ||
| + | <td width=68 valign=bottom style='width:50.95pt;background:white;padding: | ||
| + | 0cm 5.4pt 0cm 5.4pt;height:15.75pt'> | ||
| + | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;mso-line-height-alt: | ||
| + | 5.0pt'><span class=SpellE><span lang=EN-US style='font-size:10.0pt; | ||
| + | mso-fareast-font-family:"Times New Roman";color:black'>shiA</span></span><i><span | ||
| + | lang=EN-US style='font-size:10.0pt;mso-fareast-font-family:"Times New Roman"; | ||
| + | color:black'><o:p></o:p></span></i></p> | ||
| + | </td> | ||
| + | <td width=74 valign=bottom style='width:55.25pt;background:white;padding: | ||
| + | 0cm 5.4pt 0cm 5.4pt;height:15.75pt'> | ||
| + | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;mso-line-height-alt: | ||
| + | 5.0pt'><i><span lang=EN-US style='font-size:10.0pt;mso-fareast-font-family: | ||
| + | "Times New Roman";color:black'>P. <span class=SpellE>putida</span></span></i><span | ||
| + | lang=EN-US style='font-size:10.0pt;mso-fareast-font-family:"Times New Roman"; | ||
| + | color:black'><o:p></o:p></span></p> | ||
| + | </td> | ||
| + | <td width=275 valign=bottom style='width:205.9pt;border:none;border-right: | ||
| + | solid black 1.0pt;background:white;padding:0cm 5.4pt 0cm 5.4pt;height:15.75pt'> | ||
| + | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;mso-line-height-alt: | ||
| + | 5.0pt'><span lang=EN-US style='font-size:10.0pt;mso-fareast-font-family:"Times New Roman"; | ||
| + | color:black'>Removed<o:p></o:p></span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | <tr style='mso-yfti-lastrow:yes;height:15.75pt'> | ||
| + | <td width=216 valign=bottom style='width:162.0pt;border-top:none;border-left: | ||
| + | solid black 1.0pt;border-bottom:solid black 1.0pt;border-right:none; | ||
| + | background:white;padding:0cm 5.4pt 0cm 5.4pt;height:15.75pt'> | ||
| + | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;mso-line-height-alt: | ||
| + | 5.0pt'><span class=SpellE><span lang=EN-US style='font-size:10.0pt; | ||
| + | mso-fareast-font-family:"Times New Roman";color:black'>Cinnamic</span></span><span | ||
| + | lang=EN-US style='font-size:10.0pt;mso-fareast-font-family:"Times New Roman"; | ||
| + | color:black'> acid transporter<o:p></o:p></span></p> | ||
| + | </td> | ||
| + | <td width=68 valign=bottom style='width:50.95pt;border:none;border-bottom: | ||
| + | solid black 1.0pt;background:white;padding:0cm 5.4pt 0cm 5.4pt;height:15.75pt'> | ||
| + | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;mso-line-height-alt: | ||
| + | 5.0pt'><span lang=EN-US style='font-size:10.0pt;mso-fareast-font-family:"Times New Roman"; | ||
| + | color:black'> <o:p></o:p></span></p> | ||
| + | </td> | ||
| + | <td width=74 valign=bottom style='width:55.25pt;border:none;border-bottom: | ||
| + | solid black 1.0pt;background:white;padding:0cm 5.4pt 0cm 5.4pt;height:15.75pt'> | ||
| + | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;mso-line-height-alt: | ||
| + | 5.0pt'><span lang=EN-US style='font-size:10.0pt;mso-fareast-font-family:"Times New Roman"; | ||
| + | color:black'> <o:p></o:p></span></p> | ||
| + | </td> | ||
| + | <td width=275 valign=bottom style='width:205.9pt;border-top:none;border-left: | ||
| + | none;border-bottom:solid black 1.0pt;border-right:solid black 1.0pt; | ||
| + | background:white;padding:0cm 5.4pt 0cm 5.4pt;height:15.75pt'> | ||
| + | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;mso-line-height-alt: | ||
| + | 5.0pt'><span lang=EN-US style='font-size:10.0pt;mso-fareast-font-family:"Times New Roman"; | ||
| + | color:black'>Removed</span></p> | ||
| + | </td> | ||
| + | </tr> | ||
| + | </table> | ||
| + | </center> | ||
<center> | <center> | ||
Revision as of 22:21, 18 October 2012
Genetics
We plan to use a single metabolic pathway for the conversion of glucose to aromatic chemicals. We are currently engineering a Pseudomonas putida strain containing a “switch” system enabling the manufacture of either shikimic acid (SA), cinnamic acid (CA) as a major product. Future embodiments of this strain will contain additional directing mechanisms whereby a multitude of derivative chemicals can be favored.
Our metabolic engineering strategy is based on the invention of Dr. John Frost (US Patent No. 5168056, 6613552). The enzymatic activities leading to the production of SA will be enhanced by the addition of extra-chromosomal copies of endogenous genes modified such that expression is constitutive . Where necessary, these genes will be further modified through site-directed mutagenesis to abolish feedback regulation. To enhance the yield of our desired products, two additional categories of changes will be made to our host strain: (1) Enzymatic activities which direct metabolites away from the desired pathway will be abolished by removing the underlying endogenous genes and (2) transporter proteins responsible for re-uptake of SA or CA from the medium will be removed in the same manner.
Enzymatic activities which lead from SA to phenylalanine (Phe) will be abolished by removing the underlying endogenous genes. These endogenous genes, in addition to a codon-optimized Rhodosporidium toruloides gene encoding for a phenylalanine ammonia-lysase (PAL) enzyme, will be re-introduced at an extra-chromosomal locus as modified genes containing inducible promoter/repressor “switches” allowing us to direct metabolism toward CA at will. The “switch” mechanism will require “cis” DNA elements immediately upstream of each enzyme’s coding sequence (CDS). In addition, each regulatory system (ie. induction, repression) will require a constitutively-regulated “trans” element encoded at a different locus which produces the protein component.
|
Part name |
Gene/Locus |
Source |
Note |
|
Constitutive promoter |
n/a |
Synthetic |
General part |
|
Inducible promoter (cis) |
n/a |
Synthetic |
General part |
|
Inducible promoter (trans) |
n/a |
Synthetic |
General part |
|
Inducible repressor (cis) |
n/a |
Synthetic |
General part |
|
Inducible repressor (trans) |
n/a |
Synthetic |
General part |
|
Ribosome binding site |
n/a |
Synthetic |
General part |
|
Transcription terminator |
n/a |
Synthetic |
General part |
|
Transketolase |
PPS_3480 |
P. putida |
Constitutively upregulated |
|
DAHP synthase |
PPS_1503 |
P. putida |
Constitutively upregulated, Thr326Pro
= feedback insensitive |
|
DHQ synthase |
PPS_4923 |
P. putida |
Constitutively upregulated |
|
DHQ dehydratase |
PPS_0557 |
P. putida |
Constitutively upregulated |
|
Shikimate dehydrogenase |
PPS_0037 |
P. putida |
Constitutively upregulated |
|
Shikimate kinase |
PPS_4924 |
P. putida |
Removed and reintroduced as inducible |
|
3-phosphoshikimate 1-carboxyvinyltransferase |
PPS_1411 |
P. putida |
Removed and reintroduced as inducible |
|
Chorismate synthase |
PPS_1471 |
P. putida |
Removed and reintroduced as inducible |
|
Chorismatemutase |
PPS_1410 |
P. putida |
Removed and reintroduced as inducible |
|
Aromatic amino acid aminotransferase |
PPS_1561 |
P. putida |
Removed and reintroduced as inducible |
|
Aromatic amino acid aminotransferase |
PPS_3083 |
P. putida |
Removed and reintroduced as inducible |
|
Histidinol-phosphate aminotransferase |
PPS_0995 |
P. putida |
Removed and reintroduced as inducible |
|
Phenylalanine ammonia-lysase |
|
R. toruloides |
Introduced as inducible |
|
Shikimate kinase |
PPS_4924 |
P. putida |
Removed and reintroduced as inducible |
|
3-phosphoshikimate 1-carboxyvinyltransferase |
PPS_1411 |
P. putida |
Removed and reintroduced as inducible |
|
Chorismate synthase |
PPS_1471 |
P. putida |
Removed and reintroduced as inducible |
|
Chorismatemutase |
PPS_1410 |
P. putida |
Removed and reintroduced as inducible |
|
Aromatic amino acid aminotransferase |
PPS_1561 |
P. putida |
Removed and reintroduced as inducible |
|
Aromatic amino acid aminotransferase |
PPS_3083 |
P. putida |
Removed and reintroduced as inducible |
|
Histidinol-phosphate aminotransferase |
PPS_0995 |
P. putida |
Removed and reintroduced as inducible |
|
Pyrroloquinolinequinone |
PPS_3064 |
P. putida |
Removed |
|
Anthranilate synthase component I |
PPS_0413 |
P. putida |
Removed |
|
Anthranilate synthase component II |
PPS_0416 |
P. putida |
Removed |
|
Para-aminobenzoate synthase subunit I |
PPS_1911 |
P. putida |
Removed |
|
Shikimate transporter |
shiA |
P. putida |
Removed |
|
Cinnamic acid transporter |
|
|
Removed |
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