Part:BBa_I746909:Experience

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Applications of BBa_I746909

2018 iGEM team Linkoping Sweden

2018 iGEM team Linkoping Sweden validated this part.

The IPTG induction of BBa_I746909 was validated by expressing the protein in E. coli. Fluorescent was measured over 16 hours after IPTG induction and the bacteria was incubated at 37 ͦC. excitation at 488 nm and emission at 510 nm. fluorescent was measured. The optical density was measured at 600 nm. Different induction concentration of IPTG was used to stimulate the expression. experiments where conducted in an infinite M1000 PRO plate reader. Graphs show the mean values of fluorescents per bacteria of 6 biological replicates. The error bars represent the 95 % confidence interval. Results seem to indicate that 10 mM IPTG will give the most effective response, that the IPTG promoter is very sensitive yet somewhat leaky.

SKRIV HÄR!!!!!!!!!!

[http://2014.igem.org/Team:Imperial iGEM Team Imperial

We used this part as a template to make our own part (K1321337). We added Freiburg sites onto the existing sfGFP using overhang end primers during PCR. The part we made was sequence confirmed and showed very bright fluorescence, even when fused to our CBDs (cellulose binding domains).

[http://2014.igem.org/Team:Aachen iGEM Team Aachen

The iGEM Team Aachen used the biobrick I746909 to test their [http://2014.igem.org/Team:Aachen/Project/2D_Biosensor sensor chip technology].

Testing I746909 in sensor chips
I746909 in sensor chips induced with 2µl IPTG and measured with a plate reader. Top chip is not induced, bottom chip is induced with IPTG.

The biorbick I746909 was also used to test the measurement device [http://2014.igem.org/Team:Aachen/Project/Measurement_Device WatsOn].

Testing I746909 in WatsOn
I746909 in sensor chips induced with 2µl IPTG and measured with WatsOn. Left chip is not induced, right chip is induced with IPTG.

2015 iGEM team Bielefeld-CeBiTec

<a href="http://2015.igem.org/Team:Bielefeld-CeBiTec">2015 iGEM team Bielefeld-CeBiTec</a> used this part and improved it by addition of a designed, translation enhancing 5'-untranslated region (5'-UTR; <a href="https://parts.igem.org/Part:BBa_K1758100">BBa_K1758100</a>). You can find further information at <a href="https://parts.igem.org/Part:BBa_K1758102">BBa_K1758102</a>).

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In vivo characterization of sfGFP with and without our designed, translation enhancing 5'-untranslated region (5'-UTR; BBa_K1758100). Relative fluorescence units were normalized on OD600. Error bars represent standard deviation of triplicates. PT7-sfGFP: BBa_I746909 (this part); PT7-UTR-sfGFP: BBa_K1758102

As can be seen in the picture, the difference was observable with the naked eye as well.

Comparision of cultures expressing sfGFP with the optimized UTR and without. Left: BBa_K1758102, right: BBa_I746909 (this part)

[http://2014.igem.org/Team:Imperial iGEM Team Imperial

We used this part as a template to make our own part (K1321337). We added Freiburg sites onto the existing sfGFP using overhang end primers during PCR. The part we made was sequence confirmed and showed very bright fluorescence, even when fused to our CBDs (cellulose binding domains).

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[http://2014.igem.org/Team:Aachen iGEM Team Aachen

The iGEM Team Aachen used the biobrick I746909 to test their [http://2014.igem.org/Team:Aachen/Project/2D_Biosensor sensor chip technology].

Testing I746909 in sensor chips
I746909 in sensor chips induced with 2µl IPTG and measured with a plate reader. Top chip is not induced, bottom chip is induced with IPTG.

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The biorbick I746909 was also used to test the measurement device [http://2014.igem.org/Team:Aachen/Project/Measurement_Device WatsOn].

Testing I746909 in WatsOn
I746909 in sensor chips induced with 2µl IPTG and measured with WatsOn. Left chip is not induced, right chip is induced with IPTG.

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User Reviews

UNIQd530d02af6344eb9-partinfo-00000005-QINU

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