Team:Dalhousie/InterLab

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Interlab

Scientific discovery depends on having valid results. Without these valid results it becomes difficult to make any real conclusions or progress. One way that variability can arise between teams is through measurements, especially measurements that involve complicated machines and procedures. The InterLab study is a way to test how different measurements can be between teams while preforming the same experiment. This year the InterLab study is looking at variability in fluorescence, by having teams across the world measure the same glowing protein. More specifically, each team will look at the same six promoters and look at their affect on green fluorescent protein (GFP).

Background

Our Experiment

• We will be performing the same experiment as teams across the world. We clone genes into six sets of bacteria, each with the same GFP protein but different promoter regions and ribosome binding sites.

What are promoters?

• Promoters are sections of DNA that control the expression of genes, meaning they decide if the protein is created or not. Some people find it useful to think of promoters as switches on the assembly line, they don’t build the protein, but they need to be present for synthesis to occur. Its also important to know that not all promoters are the same, they differ in the amount of expression or synthesis they cause.

Constructs

• This experiment tested the effects of three promoters and two ribosome-binding sequences (RBS). All of the constructs used were delivered and used in plasmid form and encoded for chloramphenicol resistance. In total there were six constructs. Each had one of the following promoters j23106, j23101 and j23117. Along with one promoter, each construct also had one of the two RBS (b0034 or j364100). Every construct had the same GFP (e0040) and terminator sequence (b0012).

Experimental

• Along with the six experimental constructs, iGEM included two control constructs to verify the measurements recorded. A positive control used the same constant GFP and terminator sequence with a known promoter (j23151) and RBS (b00032). A negative control was also included to safeguard against contamination. This construct had a pTetR promoter with no downstream GFP included.

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