Difference between revisions of "Part:BBa K2826008"
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4. all samples were added to a 96 well plate, and the corresponding OD600 value and fluorescence intensity value were read out by Tecan Infinite M200. | 4. all samples were added to a 96 well plate, and the corresponding OD600 value and fluorescence intensity value were read out by Tecan Infinite M200. | ||
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| + | '''experimental result'''： | ||
[[Image:Copa.s1.png|600px|thumb|centre|Fig.1.Changes in fluorescence induced by different thickness of copper ion in Copa.s1]] | [[Image:Copa.s1.png|600px|thumb|centre|Fig.1.Changes in fluorescence induced by different thickness of copper ion in Copa.s1]] | ||
Revision as of 17:08, 17 October 2018
copper sensor1
A device to detect the copper ion
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 898
Illegal AgeI site found at 1010 - 1000COMPATIBLE WITH RFC[1000]
Usage and Biology
The device contains a copperA promoter,a riboJ,a RBS,a RFP repoter and two terminator. This device will be used to detect copper in solution. The copA promoter is active in the presence of copper. RiboJ can reliably maintain relative promoter strengths. RiboJ helps reduce the leakage of copA promoter greatly. After adding copper ions, the expression of red-fluorescent protein increased steadily.
CopA, the principal copper efflux ATPase in Escherichia coli, is induced by elevated copper in the medium.The copA promoter is Copper-responsive and regulated by CueR,a Member of the MerR Family.
Copper ion detection experiment
experimental method：
1. take 50mL centrifuge tubes and labeled them by marker pen. Each centrifuge tube was added with 2.5mL LB medium and 2.5mLSterilized water, 15 μL chloramphenicol. When measuring wastewater, replace 2.5 ml of ultrapure water with 2.5 ml of waste water.
2. Take tube inoculate 0.02OD strain (DH5a transformed with plasmid copper sensor1 or 2) in each tube, and sealed with a lid. cultured at 37 degrees on a shaker for 10 hours.
3. Take 100ul sample every two hours and store on ice.
4. all samples were added to a 96 well plate, and the corresponding OD600 value and fluorescence intensity value were read out by Tecan Infinite M200.
experimental result：
