Difference between revisions of "Part:BBa K2533018"
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<h1>'''Characterization'''</h1> | <h1>'''Characterization'''</h1> | ||
We add FLAG-tag and His-tag to help us to detect whether the protein has been expressed or not. | We add FLAG-tag and His-tag to help us to detect whether the protein has been expressed or not. | ||
| − | [[File:T--HUST-China--2018-（Dn）-（1）.png|400px|thumb|center|Figure1 | + | [[File:T--HUST-China--2018-（Dn）-（1）.png|400px|thumb|center|Figure1. RBS-ldhD-FLAG-lldP-his tag]] |
<h2> Fluorescence image</h2> | <h2> Fluorescence image</h2> | ||
To make sure that we get the target gene, we did the detection of fluorescence microscopy .And here is the result.Needless to see that the gene expressed well. The fluorescence of YFP is much more brighter than negative control.Therefore ,to some extant the expression is was successful. | To make sure that we get the target gene, we did the detection of fluorescence microscopy .And here is the result.Needless to see that the gene expressed well. The fluorescence of YFP is much more brighter than negative control.Therefore ,to some extant the expression is was successful. | ||
| − | [[File:T--HUST-China--2018-（YFP）-（1）.png|400px|thumb|center|Figure2 | + | [[File:T--HUST-China--2018-（YFP）-（1）.png|400px|thumb|center|Figure2. fluorescence microscopy]] |
<h2>lactate detect</h2> | <h2>lactate detect</h2> | ||
| − | [[File:T--HUST-China--2018-（乳酸）-（1）.png|400px|thumb|center|Figure3 | + | [[File:T--HUST-China--2018-（乳酸）-（1）.png|400px|thumb|center|Figure3. Result of lactate detecting]] |
Lactate production of engineered Synechocystis PCC6803. Comparison of WT, ldhD-lldP, ldhDc-lldP, ldhDnARSdR-lldP, ldhDARSdR-lldP. Synechocystis grew for 7 days(OD=1), then L-Rhamose(1g/L) was added. After induced 72 hours, lactate concentration in culture supernatant has shown above.Obviously ,we can see the engineer sphaerocystis produced more lactate. | Lactate production of engineered Synechocystis PCC6803. Comparison of WT, ldhD-lldP, ldhDc-lldP, ldhDnARSdR-lldP, ldhDARSdR-lldP. Synechocystis grew for 7 days(OD=1), then L-Rhamose(1g/L) was added. After induced 72 hours, lactate concentration in culture supernatant has shown above.Obviously ,we can see the engineer sphaerocystis produced more lactate. | ||
Latest revision as of 22:47, 17 October 2018
RBS-ldhDnARSdR-FLAG-RBS-lldP-his tag
Show the expression of the protein.
Characterization
We add FLAG-tag and His-tag to help us to detect whether the protein has been expressed or not.
Fluorescence image
To make sure that we get the target gene, we did the detection of fluorescence microscopy .And here is the result.Needless to see that the gene expressed well. The fluorescence of YFP is much more brighter than negative control.Therefore ,to some extant the expression is was successful.
lactate detect
Lactate production of engineered Synechocystis PCC6803. Comparison of WT, ldhD-lldP, ldhDc-lldP, ldhDnARSdR-lldP, ldhDARSdR-lldP. Synechocystis grew for 7 days(OD=1), then L-Rhamose(1g/L) was added. After induced 72 hours, lactate concentration in culture supernatant has shown above.Obviously ,we can see the engineer sphaerocystis produced more lactate.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 780
Illegal BglII site found at 1357 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 1493
Illegal AgeI site found at 1956
Illegal AgeI site found at 2394
Illegal AgeI site found at 2553
Illegal AgeI site found at 2565 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 764
Illegal BsaI.rc site found at 1067