Difference between revisions of "Team:Sydney Australia/Parts"
| Line 246: | Line 246: | ||
mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:78.9pt"> | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:78.9pt"> | ||
<p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand"><o:p> </o:p></span></p> | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand"><o:p> </o:p></span></p> | ||
| − | <p class=" | + | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand">Novel short chain insulin |
| − | + | analogue (<span class="SpellE">Winsulin</span>) with a 12AA linker <br> | |
| − | + | (-QRGGGSGGGQRR-)<o:p></o:p></span></p> | |
| − | <p class=" | + | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand"><o:p> </o:p></span></p> |
| − | + | <p class="MsoNormal" align="center" style="text-align:center"><span class="SpellE"><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand">YncM</span></span><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand"> secretion tag to | |
translocate protein into media<o:p></o:p></span></p> | translocate protein into media<o:p></o:p></span></p> | ||
| − | <p class=" | + | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand"><o:p> </o:p></span></p> |
| − | + | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand">N-terminal 6x His Tag | |
| − | + | purification<o:p></o:p></span></p> | |
| − | <p class=" | + | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand"><o:p> </o:p></span></p> |
| − | + | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand">4x GSS flexible linker<o:p></o:p></span></p> | |
| − | + | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand"><o:p> </o:p></span></p> | |
| − | <p class=" | + | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand">TEV protease cleavage site<o:p></o:p></span></p> |
| − | + | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand"><o:p> </o:p></span></p> | |
| − | + | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand">Extended ribosome binding site<o:p></o:p></span></p> | |
| − | <p class=" | + | |
| − | + | ||
| − | + | ||
<p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand"><o:p> </o:p></span></p> | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand"><o:p> </o:p></span></p> | ||
</td> | </td> | ||
| Line 271: | Line 268: | ||
mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:78.9pt"> | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:78.9pt"> | ||
<p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand"><o:p> </o:p></span></p> | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand"><o:p> </o:p></span></p> | ||
| + | <p class="MsoNormal"><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand"><o:p> </o:p></span></p> | ||
<p class="MsoNormal"><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand">We | <p class="MsoNormal"><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand">We | ||
demonstrated that it works:<o:p></o:p></span></p> | demonstrated that it works:<o:p></o:p></span></p> | ||
| − | <p class="MsoListParagraphCxSpFirst" style="text-indent: | + | <p class="MsoListParagraphCxSpFirst" style="text-indent:-00.0pt;mso-list:l10 level1 lfo19"><!--[if !supportLists]--><span lang="EN-US" style="font-size:15.0pt;font-family:Symbol;mso-fareast-font-family: |
Symbol;mso-bidi-font-family:Symbol"><span style="mso-list:Ignore">·<span style="font:7.0pt "Times New Roman""> </span></span></span><!--[endif]--><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand">We tested it in | Symbol;mso-bidi-font-family:Symbol"><span style="mso-list:Ignore">·<span style="font:7.0pt "Times New Roman""> </span></span></span><!--[endif]--><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand">We tested it in | ||
insulin-sensitive cell lines and saw an increase in glycogen synthesis and | insulin-sensitive cell lines and saw an increase in glycogen synthesis and | ||
| Line 279: | Line 277: | ||
<!--[if !supportLineBreakNewLine]--><br style="mso-special-character:line-break"> | <!--[if !supportLineBreakNewLine]--><br style="mso-special-character:line-break"> | ||
<!--[endif]--><o:p></o:p></span></p> | <!--[endif]--><o:p></o:p></span></p> | ||
| − | <p class="MsoListParagraphCxSpMiddle" style="text-indent: | + | <p class="MsoListParagraphCxSpMiddle" style="text-indent:-00.0pt;mso-list:l10 level1 lfo19"><!--[if !supportLists]--><span lang="EN-US" style="font-size:15.0pt;font-family:Symbol;mso-fareast-font-family: |
Symbol;mso-bidi-font-family:Symbol"><span style="mso-list:Ignore">·<span style="font:7.0pt "Times New Roman""> </span></span></span><!--[endif]--><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand">We showed via an | Symbol;mso-bidi-font-family:Symbol"><span style="mso-list:Ignore">·<span style="font:7.0pt "Times New Roman""> </span></span></span><!--[endif]--><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand">We showed via an | ||
ELISA assay that whilst the cell lysate did not contain <span class="SpellE">Winsulin</span>, | ELISA assay that whilst the cell lysate did not contain <span class="SpellE">Winsulin</span>, | ||
| Line 312: | Line 310: | ||
<p class="MsoNormal"><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand"><o:p> </o:p></span></p> | <p class="MsoNormal"><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand"><o:p> </o:p></span></p> | ||
<p class="MsoNormal"><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand"><o:p> </o:p></span></p> | <p class="MsoNormal"><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand"><o:p> </o:p></span></p> | ||
| − | <p class=" | + | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand">Human proinsulin<o:p></o:p></span></p> |
| − | + | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand"><o:p> </o:p></span></p> | |
| − | <p class=" | + | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand">N-terminal <span class="SpellE">Ecotin</span> |
| − | + | tag for targeting proteins to the periplasm<o:p></o:p></span></p> | |
| − | <p class=" | + | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand"><o:p> </o:p></span></p> |
| − | + | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand">N-terminal 6x His tag<o:p></o:p></span></p> | |
| − | + | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand"><o:p> </o:p></span></p> | |
| − | <p class=" | + | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand">4x GGS flexible linker<o:p></o:p></span></p> |
| − | + | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand"><o:p> </o:p></span></p> | |
| − | + | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand">trypsin protease cleavage site | |
| − | <p class=" | + | for His-tag and C-peptide cleavage<o:p></o:p></span></p> |
| − | + | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand"><o:p> </o:p></span></p> | |
| − | + | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand">Extended ribosomal binding | |
| − | <p class=" | + | site<o:p></o:p></span></p> |
| − | + | ||
| − | + | ||
<p class="MsoNormal"><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand"><o:p> </o:p></span></p> | <p class="MsoNormal"><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand"><o:p> </o:p></span></p> | ||
</td> | </td> | ||
| Line 340: | Line 336: | ||
improved previous parts:<o:p></o:p></span></p> | improved previous parts:<o:p></o:p></span></p> | ||
<p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand"><a href="http://parts.igem.org/Part:BBa_M39904">BBa_M39904</a> and <a href="http://parts.igem.org/Part:BBa_M1877">BBa_M1877</a> by:<o:p></o:p></span></p> | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand"><a href="http://parts.igem.org/Part:BBa_M39904">BBa_M39904</a> and <a href="http://parts.igem.org/Part:BBa_M1877">BBa_M1877</a> by:<o:p></o:p></span></p> | ||
| − | <p class=" | + | <p class="MsoNormal"><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand"><o:p> </o:p></span></p> |
| − | + | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand">completing their sequence<o:p></o:p></span></p> | |
| − | + | <p class="MsoNormal"><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand"><o:p> </o:p></span></p> | |
| − | <p class=" | + | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand">Adding a periplasmic |
| − | + | transporter tag (for more efficient folding in an oxidative environment)<o:p></o:p></span></p> | |
| − | + | <p class="MsoNormal"><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand"><o:p> </o:p></span></p> | |
| − | + | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand">Adding an efficient ribosomal binding | |
| − | <p class=" | + | site<o:p></o:p></span></p> |
| − | + | <p class="MsoNormal"><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand"><o:p> </o:p></span></p> | |
| − | + | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand">Adding an N-terminal His tag | |
| − | <p class=" | + | for purification<o:p></o:p></span></p> |
| − | + | ||
| − | + | ||
<p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand"><o:p> </o:p></span></p> | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand"><o:p> </o:p></span></p> | ||
<p class="MsoNormal"><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand">We | <p class="MsoNormal"><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand">We | ||
demonstrated that it works:<o:p></o:p></span></p> | demonstrated that it works:<o:p></o:p></span></p> | ||
| − | <p class="MsoListParagraph" style="text-indent: | + | <p class="MsoListParagraph" style="text-indent:-00.0pt;mso-list:l12 level1 lfo22"><!--[if !supportLists]--><span lang="EN-US" style="font-size:15.0pt;font-family:Symbol;mso-fareast-font-family: |
Symbol;mso-bidi-font-family:Symbol"><span style="mso-list:Ignore">·<span style="font:7.0pt "Times New Roman""> </span></span></span><!--[endif]--><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand">By testing it | Symbol;mso-bidi-font-family:Symbol"><span style="mso-list:Ignore">·<span style="font:7.0pt "Times New Roman""> </span></span></span><!--[endif]--><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand">By testing it | ||
insulin-sensitive cell lines and saw an increase in glycogen synthesis and | insulin-sensitive cell lines and saw an increase in glycogen synthesis and | ||
| Line 391: | Line 385: | ||
mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt; | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt; | ||
mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:42.15pt"> | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:42.15pt"> | ||
| − | <p class=" | + | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand">Human proinsulin<o:p></o:p></span></p> |
| − | + | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand"><o:p> </o:p></span></p> | |
| − | <p class=" | + | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand">N-terminal 6x His tag<o:p></o:p></span></p> |
| − | + | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand"><o:p> </o:p></span></p> | |
| − | + | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand">4x GGS flexible linker<o:p></o:p></span></p> | |
| − | <p class=" | + | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand"><o:p> </o:p></span></p> |
| − | + | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand">Trypsin protease cleavage site | |
| − | <p class=" | + | for His-tag and C-peptide cleavage<o:p></o:p></span></p> |
| − | + | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand"><o:p> </o:p></span></p> | |
| − | + | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand">Extended ribosomal binding | |
| − | <p class=" | + | site<o:p></o:p></span></p> |
| − | + | ||
| − | + | ||
</td> | </td> | ||
<td width="41%" valign="top" style="width:41.56%;border-top:none;border-left: | <td width="41%" valign="top" style="width:41.56%;border-top:none;border-left: | ||
| Line 413: | Line 405: | ||
improved previous parts:<o:p></o:p></span></p> | improved previous parts:<o:p></o:p></span></p> | ||
<p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand"><a href="http://parts.igem.org/Part:BBa_M39904">BBa_M39904</a> and <a href="http://parts.igem.org/Part:BBa_M1877">BBa_M1877</a> by:<o:p></o:p></span></p> | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand"><a href="http://parts.igem.org/Part:BBa_M39904">BBa_M39904</a> and <a href="http://parts.igem.org/Part:BBa_M1877">BBa_M1877</a> by:<o:p></o:p></span></p> | ||
| − | <p class="MsoListParagraphCxSpFirst" style="text-indent: | + | <p class="MsoListParagraphCxSpFirst" style="text-indent:-00.0pt;mso-list:l12 level1 lfo22"><!--[if !supportLists]--><span lang="EN-US" style="font-size:15.0pt;font-family:Symbol;mso-fareast-font-family: |
Symbol;mso-bidi-font-family:Symbol"><span style="mso-list:Ignore">·<span style="font:7.0pt "Times New Roman""> </span></span></span><!--[endif]--><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand">completing their | Symbol;mso-bidi-font-family:Symbol"><span style="mso-list:Ignore">·<span style="font:7.0pt "Times New Roman""> </span></span></span><!--[endif]--><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand">completing their | ||
sequence<o:p></o:p></span></p> | sequence<o:p></o:p></span></p> | ||
| − | <p class="MsoListParagraphCxSpMiddle" style="text-indent: | + | <p class="MsoListParagraphCxSpMiddle" style="text-indent:-00.0pt;mso-list:l12 level1 lfo22"><!--[if !supportLists]--><span lang="EN-US" style="font-size:15.0pt;font-family:Symbol;mso-fareast-font-family: |
Symbol;mso-bidi-font-family:Symbol"><span style="mso-list:Ignore">·<span style="font:7.0pt "Times New Roman""> </span></span></span><!--[endif]--><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand">Adding an efficient | Symbol;mso-bidi-font-family:Symbol"><span style="mso-list:Ignore">·<span style="font:7.0pt "Times New Roman""> </span></span></span><!--[endif]--><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand">Adding an efficient | ||
ribosomal binding site<o:p></o:p></span></p> | ribosomal binding site<o:p></o:p></span></p> | ||
| − | <p class="MsoListParagraphCxSpMiddle" style="text-indent: | + | <p class="MsoListParagraphCxSpMiddle" style="text-indent:-00.0pt;mso-list:l12 level1 lfo22"><!--[if !supportLists]--><span lang="EN-US" style="font-size:15.0pt;font-family:Symbol;mso-fareast-font-family: |
Symbol;mso-bidi-font-family:Symbol"><span style="mso-list:Ignore">·<span style="font:7.0pt "Times New Roman""> </span></span></span><!--[endif]--><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand">Adding an | Symbol;mso-bidi-font-family:Symbol"><span style="mso-list:Ignore">·<span style="font:7.0pt "Times New Roman""> </span></span></span><!--[endif]--><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand">Adding an | ||
N-terminal His tag for purification<o:p></o:p></span></p> | N-terminal His tag for purification<o:p></o:p></span></p> | ||
| Line 426: | Line 418: | ||
<p class="MsoListParagraphCxSpMiddle"><span lang="EN-US" style="font-size:15.0pt; | <p class="MsoListParagraphCxSpMiddle"><span lang="EN-US" style="font-size:15.0pt; | ||
font-family:Quicksand">We demonstrated that it works:<o:p></o:p></span></p> | font-family:Quicksand">We demonstrated that it works:<o:p></o:p></span></p> | ||
| − | <p class="MsoListParagraphCxSpLast" style="text-indent: | + | <p class="MsoListParagraphCxSpLast" style="text-indent:-00.0pt;mso-list:l12 level1 lfo22"><!--[if !supportLists]--><span lang="EN-US" style="font-size:15.0pt;font-family:Symbol;mso-fareast-font-family: |
Symbol;mso-bidi-font-family:Symbol"><span style="mso-list:Ignore">·<span style="font:7.0pt "Times New Roman""> </span></span></span><!--[endif]--><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand">By testing it insulin-sensitive | Symbol;mso-bidi-font-family:Symbol"><span style="mso-list:Ignore">·<span style="font:7.0pt "Times New Roman""> </span></span></span><!--[endif]--><span lang="EN-US" style="font-size:15.0pt;font-family:Quicksand">By testing it insulin-sensitive | ||
cell lines and saw an increase in glycogen synthesis and glucose oxidation | cell lines and saw an increase in glycogen synthesis and glucose oxidation | ||
| Line 436: | Line 428: | ||
</tr> | </tr> | ||
</tbody></table> | </tbody></table> | ||
| − | |||
</div> | </div> | ||
</div> | </div> | ||
| Line 478: | Line 469: | ||
none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt; | none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt; | ||
padding:0cm 5.4pt 0cm 5.4pt;height:11.35pt"> | padding:0cm 5.4pt 0cm 5.4pt;height:11.35pt"> | ||
| + | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:14.0pt;font-family:Quicksand"><o:p> </o:p></span></p> | ||
| + | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:14.0pt;font-family:Quicksand"><o:p> </o:p></span></p> | ||
| + | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:14.0pt;font-family:Quicksand"><o:p> </o:p></span></p> | ||
| + | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:14.0pt;font-family:Quicksand"><o:p> </o:p></span></p> | ||
<p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:14.0pt;font-family:Quicksand">Cytoplasmic-<span class="SpellE">Winsulin</span><o:p></o:p></span></p> | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:14.0pt;font-family:Quicksand">Cytoplasmic-<span class="SpellE">Winsulin</span><o:p></o:p></span></p> | ||
<p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:14.0pt;font-family:Quicksand"><a href="http://parts.igem.org/Part:BBa_K2417003">BBa_K2417003</a><o:p></o:p></span></p> | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:14.0pt;font-family:Quicksand"><a href="http://parts.igem.org/Part:BBa_K2417003">BBa_K2417003</a><o:p></o:p></span></p> | ||
| Line 496: | Line 491: | ||
mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:11.35pt"> | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:11.35pt"> | ||
<p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:14.0pt;font-family:Quicksand"><o:p> </o:p></span></p> | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:14.0pt;font-family:Quicksand"><o:p> </o:p></span></p> | ||
| − | <p class=" | + | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:14.0pt;font-family:Quicksand">Novel single chain insulin |
| − | + | analogue (<span class="SpellE">Winsulin</span>) with a 12AA linker (QRGGGSGGGQRR)<o:p></o:p></span></p> | |
| − | + | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:14.0pt;font-family:Quicksand"><o:p> </o:p></span></p> | |
| − | + | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:14.0pt;font-family:Quicksand">N-terminal 6x His Tag<o:p></o:p></span></p> | |
| − | <p class=" | + | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:14.0pt;font-family:Quicksand"><o:p> </o:p></span></p> |
| − | + | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:14.0pt;font-family:Quicksand">4x GSS flexible linker<o:p></o:p></span></p> | |
| − | + | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:14.0pt;font-family:Quicksand"><o:p> </o:p></span></p> | |
| − | <p class=" | + | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:14.0pt;font-family:Quicksand">TEV protease cleavage site<o:p></o:p></span></p> |
| − | + | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:14.0pt;font-family:Quicksand"><o:p> </o:p></span></p> | |
| − | + | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:14.0pt;font-family:Quicksand">Extended ribosome binding site<o:p></o:p></span></p> | |
| − | <p class=" | + | |
| − | + | ||
| − | + | ||
| − | <p class=" | + | |
| − | + | ||
| − | + | ||
<p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:14.0pt;font-family:Quicksand"><o:p> </o:p></span></p> | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:14.0pt;font-family:Quicksand"><o:p> </o:p></span></p> | ||
</td> | </td> | ||
| Line 520: | Line 509: | ||
<p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:14.0pt;font-family:Quicksand"><o:p> </o:p></span></p> | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:14.0pt;font-family:Quicksand"><o:p> </o:p></span></p> | ||
<p class="MsoNormal"><span lang="EN-US" style="font-size:14.0pt;font-family:Quicksand"><o:p> </o:p></span></p> | <p class="MsoNormal"><span lang="EN-US" style="font-size:14.0pt;font-family:Quicksand"><o:p> </o:p></span></p> | ||
| − | <p class="MsoListParagraph" style="text-indent: | + | <p class="MsoListParagraph" style="text-indent:-00.0pt;mso-list:l12 level1 lfo22"><!--[if !supportLists]--><span lang="EN-US" style="font-size:14.0pt;font-family:Symbol;mso-fareast-font-family: |
Symbol;mso-bidi-font-family:Symbol"><span style="mso-list:Ignore">·<span style="font:7.0pt "Times New Roman""> </span></span></span><!--[endif]--><span lang="EN-US" style="font-size:14.0pt;font-family:Quicksand">We validated our | Symbol;mso-bidi-font-family:Symbol"><span style="mso-list:Ignore">·<span style="font:7.0pt "Times New Roman""> </span></span></span><!--[endif]--><span lang="EN-US" style="font-size:14.0pt;font-family:Quicksand">We validated our | ||
parts by showing that they work in producing insulin. We performed an ELISA | parts by showing that they work in producing insulin. We performed an ELISA | ||
| Line 545: | Line 534: | ||
<div class="box3"></div> | <div class="box3"></div> | ||
<div class="row" style="height:2vw;"></div> | <div class="row" style="height:2vw;"></div> | ||
| + | <div align="center"> | ||
<div align="center"> | <div align="center"> | ||
| Line 577: | Line 567: | ||
border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt; | border-top:none;mso-border-top-alt:solid windowtext .5pt;mso-border-alt:solid windowtext .5pt; | ||
padding:0cm 5.4pt 0cm 5.4pt;height:35.15pt"> | padding:0cm 5.4pt 0cm 5.4pt;height:35.15pt"> | ||
| + | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:14.0pt;font-family:Quicksand"><o:p> </o:p></span></p> | ||
| + | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:14.0pt;font-family:Quicksand"><o:p> </o:p></span></p> | ||
| + | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:14.0pt;font-family:Quicksand"><o:p> </o:p></span></p> | ||
| + | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:14.0pt;font-family:Quicksand"><o:p> </o:p></span></p> | ||
| + | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:14.0pt;font-family:Quicksand"><o:p> </o:p></span></p> | ||
| + | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:14.0pt;font-family:Quicksand"><o:p> </o:p></span></p> | ||
| + | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:14.0pt;font-family:Quicksand"><o:p> </o:p></span></p> | ||
| + | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:14.0pt;font-family:Quicksand"><o:p> </o:p></span></p> | ||
<p class="MsoNormal" align="center" style="text-align:center"><span class="SpellE"><span lang="EN-US" style="font-size:14.0pt;font-family:Quicksand">Ecotin-Winsulin</span></span><span lang="EN-US" style="font-size:14.0pt;font-family:Quicksand"><o:p></o:p></span></p> | <p class="MsoNormal" align="center" style="text-align:center"><span class="SpellE"><span lang="EN-US" style="font-size:14.0pt;font-family:Quicksand">Ecotin-Winsulin</span></span><span lang="EN-US" style="font-size:14.0pt;font-family:Quicksand"><o:p></o:p></span></p> | ||
<p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:14.0pt;font-family:Quicksand"><a href="http://parts.igem.org/Part:BBa_K2417002">BBa_K2417002</a><o:p></o:p></span></p> | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:14.0pt;font-family:Quicksand"><a href="http://parts.igem.org/Part:BBa_K2417002">BBa_K2417002</a><o:p></o:p></span></p> | ||
| Line 593: | Line 591: | ||
mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt; | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt; | ||
mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:35.15pt"> | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:35.15pt"> | ||
| − | <p class=" | + | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:14.0pt;font-family:Quicksand"><o:p> </o:p></span></p> |
| − | + | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:14.0pt;font-family:Quicksand">Novel single chain insulin | |
| − | + | analogue (<span class="SpellE">Winsulin</span>) with a 12AA linker (QRGGGSGGGQRR)<o:p></o:p></span></p> | |
| − | <p class=" | + | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:14.0pt;font-family:Quicksand"><o:p> </o:p></span></p> |
| − | + | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:14.0pt;font-family:Quicksand">N-terminal <span class="SpellE">Ecotin</span> | |
| − | <p class=" | + | tag for targeting proteins to the periplasm<o:p></o:p></span></p> |
| − | + | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:14.0pt;font-family:Quicksand"><o:p> </o:p></span></p> | |
| − | + | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:14.0pt;font-family:Quicksand">N-terminal 6x His Tag<o:p></o:p></span></p> | |
| − | <p class=" | + | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:14.0pt;font-family:Quicksand"><o:p> </o:p></span></p> |
| − | + | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:14.0pt;font-family:Quicksand">4x GSS flexible linker<o:p></o:p></span></p> | |
| − | + | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:14.0pt;font-family:Quicksand"><o:p> </o:p></span></p> | |
| − | <p class=" | + | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:14.0pt;font-family:Quicksand">TEV protease cleavage site<o:p></o:p></span></p> |
| − | + | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:14.0pt;font-family:Quicksand"><o:p> </o:p></span></p> | |
| − | + | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:14.0pt;font-family:Quicksand">Extended ribosome binding site<o:p></o:p></span></p> | |
| − | <p class=" | + | |
| − | + | ||
| − | + | ||
</td> | </td> | ||
<td width="41%" rowspan="2" style="width:41.1%;border-top:none;border-left: | <td width="41%" rowspan="2" style="width:41.1%;border-top:none;border-left: | ||
| Line 615: | Line 610: | ||
mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt; | mso-border-top-alt:solid windowtext .5pt;mso-border-left-alt:solid windowtext .5pt; | ||
mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:35.15pt"> | mso-border-alt:solid windowtext .5pt;padding:0cm 5.4pt 0cm 5.4pt;height:35.15pt"> | ||
| + | <p class="MsoNormal"><span lang="EN-US" style="font-size:14.0pt;font-family:Quicksand"><o:p> </o:p></span></p> | ||
| + | <p class="MsoNormal"><span lang="EN-US" style="font-size:14.0pt;font-family:Quicksand"><o:p> </o:p></span></p> | ||
| + | <p class="MsoNormal"><span lang="EN-US" style="font-size:14.0pt;font-family:Quicksand"><o:p> </o:p></span></p> | ||
| + | <p class="MsoNormal"><span lang="EN-US" style="font-size:14.0pt;font-family:Quicksand"><o:p> </o:p></span></p> | ||
| + | <p class="MsoNormal"><span lang="EN-US" style="font-size:14.0pt;font-family:Quicksand"><o:p> </o:p></span></p> | ||
| + | <p class="MsoNormal"><span lang="EN-US" style="font-size:14.0pt;font-family:Quicksand"><o:p> </o:p></span></p> | ||
| + | <p class="MsoNormal"><span lang="EN-US" style="font-size:14.0pt;font-family:Quicksand"><o:p> </o:p></span></p> | ||
<p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:14.0pt;font-family:Quicksand">We submitted this DNA part to | <p class="MsoNormal" align="center" style="text-align:center"><span lang="EN-US" style="font-size:14.0pt;font-family:Quicksand">We submitted this DNA part to | ||
the registry, for other teams to use in the future<o:p></o:p></span></p> | the registry, for other teams to use in the future<o:p></o:p></span></p> | ||
| Line 632: | Line 634: | ||
</tr> | </tr> | ||
</tbody></table> | </tbody></table> | ||
| + | |||
| + | </div> | ||
</div> | </div> | ||
Revision as of 10:30, 30 October 2017
GOLD
|
PART NAME / NO. |
SOURCE |
KEY FEATURES |
WHY IS IT GOLD? |
|
YncM-Winsulin |
Bacillus subtilis (YncM secretion tag) |
Novel short chain insulin
analogue (Winsulin) with a 12AA linker YncM secretion tag to
translocate protein into media N-terminal 6x His Tag
purification 4x GSS flexible linker TEV protease cleavage site Extended ribosome binding site |
We
demonstrated that it works: · We tested it in
insulin-sensitive cell lines and saw an increase in glycogen synthesis and
glucose oxidation above basal levels, proving its bioactivity · We showed via an
ELISA assay that whilst the cell lysate did not contain Winsulin,
the media in which the cells were growing did. Therefore
the YncM secretion tag worked as expected The
results for these assays can be found here |
|
Ecotin-Proinsulin |
E. Coli (Ecotin
tag) |
Human proinsulin N-terminal Ecotin
tag for targeting proteins to the periplasm N-terminal 6x His tag 4x GGS flexible linker trypsin protease cleavage site
for His-tag and C-peptide cleavage Extended ribosomal binding
site |
We
improved previous parts: BBa_M39904 and BBa_M1877 by: completing their sequence Adding a periplasmic
transporter tag (for more efficient folding in an oxidative environment) Adding an efficient ribosomal binding
site Adding an N-terminal His tag
for purification We
demonstrated that it works: · By testing it
insulin-sensitive cell lines and saw an increase in glycogen synthesis and
glucose oxidation above basal levels, therefore proving its bioactivity The
results for these assays can be found here |
|
Human (Human
proinsulin) |
|||
|
Cytoplasmic-Proinsulin BBa_K2417001 |
Human (Human
proinsulin) |
Human proinsulin N-terminal 6x His tag 4x GGS flexible linker Trypsin protease cleavage site
for His-tag and C-peptide cleavage Extended ribosomal binding
site |
We
improved previous parts: BBa_M39904 and BBa_M1877 by: · completing their
sequence · Adding an efficient
ribosomal binding site · Adding an
N-terminal His tag for purification We demonstrated that it works: · By testing it insulin-sensitive
cell lines and saw an increase in glycogen synthesis and glucose oxidation
above basal levels, therefore proving its bioactivity The
results for these assays can be found here |
SILVER
|
PART NAME / NO. |
SOURCE |
KEY FEATURES |
WHY IS IT SILVER? |
|
Cytoplasmic-Winsulin |
N/A – Winsulin is self designed from literature and mammalian insulins |
Novel single chain insulin
analogue (Winsulin) with a 12AA linker (QRGGGSGGGQRR) N-terminal 6x His Tag 4x GSS flexible linker TEV protease cleavage site Extended ribosome binding site |
· We validated our
parts by showing that they work in producing insulin. We performed an ELISA
assay testing for properly folded insulin, and detected the presence of
Cytoplasmic Winsulin. The
results for the assay can be found here |
BRONZE
|
PART NAME / NO. |
SOURCE |
KEY FEATURES |
WHY IS IT BRONZE? |
|
Ecotin-Winsulin |
E. Coli (Ecotin
Tag) |
Novel single chain insulin
analogue (Winsulin) with a 12AA linker (QRGGGSGGGQRR) N-terminal Ecotin
tag for targeting proteins to the periplasm N-terminal 6x His Tag 4x GSS flexible linker TEV protease cleavage site Extended ribosome binding site |
We submitted this DNA part to
the registry, for other teams to use in the future |
|
N/A – Winsulin is self designed from literature and mammalian insulins |
