Team:HokkaidoU Japan/Proof
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Team:HokkaidoU Japan
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We made a functonal unit to regulate gene expression. BBa_K2015012 codes constitutive promoter (BBa_J23101), RBS (BBa_B0032), LacI (BBa_C0012), dT (BBa_B0015), PLac (BBa_R0011) and RBS (BBa_B0034). To insert mRFP at the downstream we identified this construct could regulate the expression (Fig. 1).
And also we conducted SDS-PAGE with (BBa_K2015012 + BBa_K2015008 / BBa_K2015012 + BBa_K2015009) on pSB1C3 of E. coli (DH5α). Let us explain the 3 parts below. BBa_K2015012 part contains lacI expression unit and plac + RBS. The sequences of downstream can be induced strictly by IPTG. BBa_K2015008 consists Self Assembling Regions (SAR, RADA16-I) and mutated GFP. And BBa_K2015009 this part consists Self Assembling Region (SAR, P11-4) and mutated GFP.
The results are shown the below pictures.The fluorescence of GFPs was not detected any condition
Fig. 2. BBa_K2015012-BBa_K2015009
BBa_K2015012-BBa_K2015009 on pSB1C3 vector were induced by IPTG and incubated at 25°C for 24 h (left)
and at 37°C for 16 h (right). (-): not induced, (+): 0.5 mM IPTG and (++): 1.0 mM IPTG
.
We made a functonal unit to regulate gene expression. BBa_K2015012 codes constitutive promoter (BBa_J23101), RBS (BBa_B0032), LacI (BBa_C0012), dT (BBa_B0015), PLac (BBa_R0011) and RBS (BBa_B0034). To insert mRFP at the downstream we identified this construct could regulate the expression (Fig. 1).
 |
| Fig. 1. Change of expression by IPTG induction left: not induced, right: induced |
And also we conducted SDS-PAGE with (BBa_K2015012 + BBa_K2015008 / BBa_K2015012 + BBa_K2015009) on pSB1C3 of E. coli (DH5α). Let us explain the 3 parts below. BBa_K2015012 part contains lacI expression unit and plac + RBS. The sequences of downstream can be induced strictly by IPTG. BBa_K2015008 consists Self Assembling Regions (SAR, RADA16-I) and mutated GFP. And BBa_K2015009 this part consists Self Assembling Region (SAR, P11-4) and mutated GFP.
The results are shown the below pictures.The fluorescence of GFPs was not detected any condition
 |
| Fig. 1. Result of IPTG induction BBa_K2015012-BBa_K2015008 on pSB1C3 vector were induced by IPTG and incubated at 37°C. (-): not induced, (+): 0.5 mM IPTG and (++): 1.0 mM IPTG |
|
|
BBa_K2015012-BBa_K2015009 on pSB1C3 vector were induced by IPTG and incubated at 25°C for 24 h (left)
and at 37°C for 16 h (right). (-): not induced, (+): 0.5 mM IPTG and (++): 1.0 mM IPTG
.
