Difference between revisions of "Team:IIT-Madras/Basic Part"
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<p> <h3>BBaK2857001</h3> | <p> <h3>BBaK2857001</h3> | ||
| − | The BBaK2857001 BioBrick contains GFP codon-optimized for use in <em>Acinetobacter baylyi</em>. The codon-optimized sequence was generated using the CUTE tool from ChassiDex. This part was cloned into the pBAV1K vector and driven by the bacteriophage T5 promoter. This pBAV1K-T5-CO-GFP construct was transformed into <em>A. baylyi</em> and fluorometry studies were carried out. It was found that the codon-optimized GFP shows considerably higher fluorescence than the pBAV1K-T5-eGFP part (BBA_K1321309), containing eGFP codon-optimized for use in <em>E. coli</em>, used as the control, thus demonstrating that CUTE functions as | + | The BBaK2857001 BioBrick contains GFP codon-optimized for use in <em>Acinetobacter baylyi</em>. The codon-optimized sequence was generated using the CUTE tool from ChassiDex. This part was cloned into the pBAV1K vector and driven by the bacteriophage T5 promoter. This pBAV1K-T5-CO-GFP construct was transformed into <em>A. baylyi</em> and fluorometry studies were carried out. It was found that the codon-optimized GFP shows considerably higher fluorescence than the pBAV1K-T5-eGFP part (BBA_K1321309), containing eGFP codon-optimized for use in <em>E. coli</em>, used as the control, thus demonstrating that CUTE functions as expected and any required gene can be codon-optimized for <em>A. baylyi</em>. |
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<center><img src="https://static.igem.org/mediawiki/parts/7/7e/GFP_correct.png"></center><br><br> | <center><img src="https://static.igem.org/mediawiki/parts/7/7e/GFP_correct.png"></center><br><br> | ||
Latest revision as of 01:56, 18 October 2018
Best Basic Part Special Prize
BBaK2857001
The BBaK2857001 BioBrick contains GFP codon-optimized for use in Acinetobacter baylyi. The codon-optimized sequence was generated using the CUTE tool from ChassiDex. This part was cloned into the pBAV1K vector and driven by the bacteriophage T5 promoter. This pBAV1K-T5-CO-GFP construct was transformed into A. baylyi and fluorometry studies were carried out. It was found that the codon-optimized GFP shows considerably higher fluorescence than the pBAV1K-T5-eGFP part (BBA_K1321309), containing eGFP codon-optimized for use in E. coli, used as the control, thus demonstrating that CUTE functions as expected and any required gene can be codon-optimized for A. baylyi.
The optimal excitation wavelength for the codon-optimized GFP was found to be 480 nm and the optimal emission wavelength was found to be 512 nm.