Composite

Part:BBa_K1891015:Experience

Designed by: Xinyu Liu   Group: iGEM16_BNU-China   (2016-10-13)


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Applications of BBa_K1891015

In order to make the P(3HB) production process more controllable, we added heat-sensitive promoter(BBa_K873002) to phaC1-A-B1 gene sequence (BBa_K934001), constructing the heat inducible parts (BBa_K1891015). 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 E.coli.


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


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


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