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Team:NTU-Taida/Result/pFadBA
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__FORCETOC__{{:Team:NTU-Taida/Templates/Header}}{{:Team:NTU-Taida/Templates/Navbar}}{{:Team:NTU-Taida/Templates/Sidebar|Title=pFadBA Promter}}{{:Team:NTU-Taida/Templates/ContentStart}} | __FORCETOC__{{:Team:NTU-Taida/Templates/Header}}{{:Team:NTU-Taida/Templates/Navbar}}{{:Team:NTU-Taida/Templates/Sidebar|Title=pFadBA Promter}}{{:Team:NTU-Taida/Templates/ContentStart}} | ||
| - | {{:Team:NTU-Taida/Templates/BSHero|Title=P<sub>fadBA</sub> Promter|Content=}} | + | {{:Team:NTU-Taida/Templates/BSHero|Title=P<sub>fadBA</sub> Promter Function|Content=}} |
| - | + | ==Method== | |
| - | + | ||
To evaluate the P<sub>fadBA</sub> promoter function, we design a P<sub>hs</sub>-mRFP reporter construct. In experimental group we add oleic acid in media and compared to control group, which is the same colony and identical amount of bacteria but without addition of oleic acid. After 5 hr induction we measure the man fluorescence intensity to get the expression level of P<sub>hs</sub> promoter in either group. | To evaluate the P<sub>fadBA</sub> promoter function, we design a P<sub>hs</sub>-mRFP reporter construct. In experimental group we add oleic acid in media and compared to control group, which is the same colony and identical amount of bacteria but without addition of oleic acid. After 5 hr induction we measure the man fluorescence intensity to get the expression level of P<sub>hs</sub> promoter in either group. | ||
| - | + | ==Protocol== | |
#10 μL bacterial culture was cultured in 5 mL LB at 37℃, with suitable concentration of antibiotics shaking for 18 hr. | #10 μL bacterial culture was cultured in 5 mL LB at 37℃, with suitable concentration of antibiotics shaking for 18 hr. | ||
#For experimental group, 480 μL of bacteria culture was transferred to 1.5 mL eppendorf tube and added with 20 μL of oleic acid and IGEPAL(detergent) in. For control group, 500 μL bacterial culture was added in 1.5 mL eppendorf tube. | #For experimental group, 480 μL of bacteria culture was transferred to 1.5 mL eppendorf tube and added with 20 μL of oleic acid and IGEPAL(detergent) in. For control group, 500 μL bacterial culture was added in 1.5 mL eppendorf tube. | ||
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#100 μL of bacterial culture was transferred tino 96-well plate. The mRFP fluorescence intensity was measured (Excitaion: 580 nm, Emmision: 610 nm). | #100 μL of bacterial culture was transferred tino 96-well plate. The mRFP fluorescence intensity was measured (Excitaion: 580 nm, Emmision: 610 nm). | ||
| - | + | ==Data== | |
[[FIle:NTU-Taida-Result-pFadBA-fig1.png|450px|thumb|center|TEST]] | [[FIle:NTU-Taida-Result-pFadBA-fig1.png|450px|thumb|center|TEST]] | ||
'''Oleic acid induction group shows higher expression of mRFP.''' | '''Oleic acid induction group shows higher expression of mRFP.''' | ||
Revision as of 18:56, 26 September 2012
pFadBA Promter
PfadBA Promter Function
Contents |
Method
To evaluate the PfadBA promoter function, we design a Phs-mRFP reporter construct. In experimental group we add oleic acid in media and compared to control group, which is the same colony and identical amount of bacteria but without addition of oleic acid. After 5 hr induction we measure the man fluorescence intensity to get the expression level of Phs promoter in either group.
Protocol
- 10 μL bacterial culture was cultured in 5 mL LB at 37℃, with suitable concentration of antibiotics shaking for 18 hr.
- For experimental group, 480 μL of bacteria culture was transferred to 1.5 mL eppendorf tube and added with 20 μL of oleic acid and IGEPAL(detergent) in. For control group, 500 μL bacterial culture was added in 1.5 mL eppendorf tube.
- The tubes were incubated at 37℃ 5 hr for induction.
- 100 μL of bacterial culture was transferred tino 96-well plate. The mRFP fluorescence intensity was measured (Excitaion: 580 nm, Emmision: 610 nm).
Data
Oleic acid induction group shows higher expression of mRFP.
