Difference between revisions of "Part:BBa K2132001"
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<span class='h3bb'>Sequence and Features</span> | <span class='h3bb'>Sequence and Features</span> | ||
<partinfo>BBa_K2132001 SequenceAndFeatures</partinfo> | <partinfo>BBa_K2132001 SequenceAndFeatures</partinfo> | ||
| + | <html> | ||
| + | <h3>Characterization</h3> | ||
| + | <p> | ||
| + | Three different experiments were conducted to characterize these biobricks: | ||
| + | <ul> | ||
| + | <li> TEM: visualization of CsgASpyCatcherHisTag mutant biofilm </li> | ||
| + | <li> Fluoresence Binding Test </li> | ||
| + | <li> TEM: visualization of binding test </li></ul> | ||
| + | </p> | ||
| − | TEM: visualization of CsgASpyCatcherHisTag mutant biofilm | + | <h4> TEM: visualization of CsgASpyCatcherHisTag mutant biofilm </h4> |
| − | < | + | <p> |
| − | + | ...... | |
| − | Fluoresence Binding Test | + | </p> |
| + | <h4> Fluoresence Binding Test </h4> | ||
| + | <p> | ||
In order to test the effect of binding between CsgASpyCatcherHisTag mutant and inorganic nanoparticles, we apply same amount of suspended QDs solution into M63 medium which has cultured biofilm for 72h. After 30-min incubation, we used PBS to mildly wash the well, and the result was consistent with our anticipation: On the left, CsgASpyCatcherHisTag mutant were induced and thus secreted biofilm, and firmly attached with QDS and thus show bright fluorescence. Therefore, we ensure the stable coordinate bonds between CsgASpyCatcherHisTag mutant and QDs can manage to prevent QDs from being taken away by liquid flow. | In order to test the effect of binding between CsgASpyCatcherHisTag mutant and inorganic nanoparticles, we apply same amount of suspended QDs solution into M63 medium which has cultured biofilm for 72h. After 30-min incubation, we used PBS to mildly wash the well, and the result was consistent with our anticipation: On the left, CsgASpyCatcherHisTag mutant were induced and thus secreted biofilm, and firmly attached with QDS and thus show bright fluorescence. Therefore, we ensure the stable coordinate bonds between CsgASpyCatcherHisTag mutant and QDs can manage to prevent QDs from being taken away by liquid flow. | ||
| − | < | + | </p> |
| − | TEM: visualization of binding test | + | <figure> |
| + | <img src="https://parts.igem.org/File:Shanghaitechchina_SCH%2BQD.tiff" width="100%" height="100%"> | ||
| + | <figcaption> | ||
| + | <b>Fig. 1</b>: ...... | ||
| + | </figcaption> | ||
| + | </figure> | ||
| + | |||
| + | |||
| + | <h4> TEM: visualization of binding test</h4> | ||
| + | <p> | ||
transmission electron microscopy(TEM) visualize the binding effect of CsgASpyCatcherHisTag mutant E.coli with AuNPs. From the picture, it shows biofilm areas are attached by QDs and thus confirm the viability of histag on CsgASpyCatcherHisTag mutant biofilm. | transmission electron microscopy(TEM) visualize the binding effect of CsgASpyCatcherHisTag mutant E.coli with AuNPs. From the picture, it shows biofilm areas are attached by QDs and thus confirm the viability of histag on CsgASpyCatcherHisTag mutant biofilm. | ||
| − | <img> | + | </p> |
| + | |||
| + | <figure> | ||
| + | <img src="https://parts.igem.org/File:Shanghaitechchina_SCH%2BQD.tiff" width="100%" height="100%"> | ||
| + | <figcaption> | ||
| + | <b>Fig. 1</b>: ...... | ||
| + | </figcaption> | ||
| + | </figure> | ||
| + | |||
| + | </html> | ||
===Functional Parameters=== | ===Functional Parameters=== | ||
<partinfo>BBa_K2132001 parameters</partinfo> | <partinfo>BBa_K2132001 parameters</partinfo> | ||
<!-- --> | <!-- --> | ||
Revision as of 12:09, 13 October 2016
CsgASpyCatcherHisTag
This is the subunit of the biofilm of E. Coli Curli system linked to SpyCatcher and HisTag. The two added parts can be used for various functions with SpyTag and other His tag-binding materials like quantum dots.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 841
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Characterization
Three different experiments were conducted to characterize these biobricks:
- TEM: visualization of CsgASpyCatcherHisTag mutant biofilm
- Fluoresence Binding Test
- TEM: visualization of binding test
TEM: visualization of CsgASpyCatcherHisTag mutant biofilm
......
Fluoresence Binding Test
In order to test the effect of binding between CsgASpyCatcherHisTag mutant and inorganic nanoparticles, we apply same amount of suspended QDs solution into M63 medium which has cultured biofilm for 72h. After 30-min incubation, we used PBS to mildly wash the well, and the result was consistent with our anticipation: On the left, CsgASpyCatcherHisTag mutant were induced and thus secreted biofilm, and firmly attached with QDS and thus show bright fluorescence. Therefore, we ensure the stable coordinate bonds between CsgASpyCatcherHisTag mutant and QDs can manage to prevent QDs from being taken away by liquid flow.
TEM: visualization of binding test
transmission electron microscopy(TEM) visualize the binding effect of CsgASpyCatcherHisTag mutant E.coli with AuNPs. From the picture, it shows biofilm areas are attached by QDs and thus confirm the viability of histag on CsgASpyCatcherHisTag mutant biofilm.