Difference between revisions of "Part:BBa K2205017"

Revision as of 13:20, 1 November 2017

Sensynova Framework (Connector 2B + amajLime (Complete) - rCell)

Description

The part BBa_K2205017 was designed as Processor unit of the Sensynova multicellular framework for biosensors development (Figure 1).

This part was made to testing the modularity of the system by replacing the sfGFP reporter module of the Sensynova platform design with the chromoprotein variants amajLime (BBa_ K1033915).

The amajLime protein is a yellow-green chromoprotein extracted from the coral Anemonia majano. It was first extracted and characterized by Matz et al. under the name amFP486 (UniProtKB/Swiss-Prot: Q9U6Y6.1 GI: 56749103 GenBank: AF168421.1) and codon optimized for E coliby Genscript. The protein has an absorption maximum at 458 nm giving it a yellow-green colour visible to the naked eye.

The part BBa_ K1033915 is under the control of our connector 2, part BBa_K2205013, meaning it is normally repressed expect in the presence of connector 2 (C12 – RHL).

Characterisation

Initial testing of this chromoprotein reporter variant was conducted by inoculating 1ml of LB containing the antibiotic Chloramphenicol with a colony from each colour proven by colony PCR and sequencing data to be correct and grown at 37° for 2 hours as well as a control of wildtype DH5α.

Cultures were then plated onto a LB agar + Chloramphenicol plate which were then innoculated in four locations with 10μl of connector 2 (C12 - Rhl) and grown overnight resulting in the picture below.

References

Matz, M., Fradkov, A., Labas, Y., Savitsky, A., Zaraisky, A., Markelov, M. and Lukyanov, S. (1999). Nature Biotechnology, 17(10), pp.969-973.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 7
Illegal NheI site found at 30
21
INCOMPATIBLE WITH RFC[21]
Illegal BamHI site found at 301
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI site found at 776

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K2205017 short</partinfo>
-Description here
+===Description===
+<p>The part BBa_K2205017 was designed as Processor unit of the Sensynova multicellular framework for biosensors development (Figure 1). </p>
-<!-- Add more about the biology of this part here
+[[File:Framework_generic.jpg|400px|]]
-===Usage and Biology===
+<p>This part was made to testing the modularity of the system by replacing the sfGFP reporter module of the Sensynova platform design with the chromoprotein variants amajLime (BBa_ K1033915). </p>
+<p> The amajLime protein is a yellow-green chromoprotein extracted from the coral Anemonia majano. It was first extracted and characterized by Matz et al. under the name amFP486 (UniProtKB/Swiss-Prot: Q9U6Y6.1 GI: 56749103 GenBank: AF168421.1) and codon optimized for E coliby Genscript. The protein has an absorption maximum at 458 nm giving it a yellow-green colour visible to the naked eye.</p>
+<p>The part BBa_ K1033915 is under the control of our connector 2, part BBa_K2205013, meaning it is normally repressed expect in the presence of connector 2 (C12 – RHL). </p>
+[[File:Framework_Yellow_Variant_Pic.jpg|400px|]]
+===Characterisation===
+<p>Initial testing of this chromoprotein reporter variant was conducted by inoculating 1ml of LB containing the antibiotic Chloramphenicol with a colony from each colour proven by colony PCR and sequencing data to be correct and grown at 37° for 2 hours as well as a control of wildtype DH5α. </p>
+<p>Cultures were then plated onto a LB agar + Chloramphenicol plate which were then innoculated in four locations with 10μl of connector 2 (C12 - Rhl) and grown overnight resulting in the picture below. </p>
+[[File:Framework_Yellow_Variant.jpg|400px|]]
+===References===
+Matz, M., Fradkov, A., Labas, Y., Savitsky, A., Zaraisky, A., Markelov, M. and Lukyanov, S. (1999). Nature Biotechnology, 17(10), pp.969-973.
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