Difference between revisions of "Part:BBa K1891014:Experience"
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===Applications of BBa_K1891014===
 
===Applications of BBa_K1891014===
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<p>In order to make the P(3HB) production process more controllable, we added arabinose inducible promoter(<html><a href='https://parts.igem.org/Part:BBa_I0500'>BBa_I0500</a></html>)to phaC1-A-B1 gene sequence (BBa_K934001), constructing the arabinose inducible parts (<html><a href='https://parts.igem.org/Part:BBa_K1891014'>BBa_K1891014</a></html>). Finally we linked the newly synthesized parts to the standard vector pSB1C3. The new plasmids can be used directly for arabinose induced expression in <em>E.coli.</em></p>
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<p>In order to make the P(3HB) production process more controllable, we added heat-sensitive promoter(<html><a href='https://parts.igem.org/Part:BBa_K873002'>BBa_K873002</a></html>) and arabinose inducible promoter(<html><a href='https://parts.igem.org/Part:BBa_I0500'>BBa_I0500</a></html>)to phaC1-A-B1 gene sequence (BBa_K934001), constructing the heat and arabinose inducible parts (<html><a href='https://parts.igem.org/Part:BBa_K1891014'>BBa_K1891014</a></html>) and (<html><a href='https://parts.igem.org/Part:BBa_K1891015'>BBa_K1891015</a></html>). Finally we linked the newly synthesized parts to the standard vector pSB1C3. The new plasmids can be used directly for heat shock and arabinose induced expression in <em>E.coli.</em></p>
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<p>We transformed the constructed vectors to TransB(DE3) for expression, and set different conditions for inducement.</p>
 
<p>We transformed the constructed vectors to TransB(DE3) for expression, and set different conditions for inducement.</p>
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  <strong>C,D</strong>  left to right: pSB1C3 empty(37℃)，pSB1C3 empty(Arabinose induced)，P(3HB) with natural promotor，P(3HB) with pBAD 1,2(37℃)，P(3HB) with pBAD 1,2(Arabinose induced)
 
  <strong>C,D</strong>  left to right: pSB1C3 empty(37℃)，pSB1C3 empty(Arabinose induced)，P(3HB) with natural promotor，P(3HB) with pBAD 1,2(37℃)，P(3HB) with pBAD 1,2(Arabinose induced)
 
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===User Reviews===
 
===User Reviews===

Revision as of 03:08, 29 October 2016


This experience page is provided so that any user may enter their experience using this part.
Please enter how you used this part and how it worked out.

Applications of BBa_K1891014

In order to make the P(3HB) production process more controllable, we added arabinose inducible promoter(BBa_I0500)to phaC1-A-B1 gene sequence (BBa_K934001), constructing the arabinose inducible parts (BBa_K1891014). Finally we linked the newly synthesized parts to the standard vector pSB1C3. The new plasmids can be used directly for arabinose induced expression in E.coli.


We transformed the constructed vectors to TransB(DE3) for expression, and set different conditions for inducement.


experimental group induce conditions
37℃ 42℃ 1 mM Arabinose
pSB1C3 empty √ √ √
P(3HB) with natural promotor √
P(3HB) with HSP 1 √ √
P(3HB) with HSP 2 √ √
P(3HB) with pBAD 1 √ √
P(3HB) with pBAD 2 √ √


The results show as follow,


A,B left to right: pSB1C3 empty(37℃)，pSB1C3 empty(42℃ induced)，P(3HB) with natural promotor，P(3HB) with HSP 1,2(37℃)，P(3HB) with HSP 1,2(42℃).
C,D left to right: pSB1C3 empty(37℃)，pSB1C3 empty(Arabinose induced)，P(3HB) with natural promotor，P(3HB) with pBAD 1,2(37℃)，P(3HB) with pBAD 1,2(Arabinose induced)


User Reviews

UNIQ50c4a264af74cb87-partinfo-00000002-QINU UNIQ50c4a264af74cb87-partinfo-00000003-QINU