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− | <p align="left" style="background-color:#ffffff;"> <br><br><br>Team TSH consisted of Emily, Helen and Sarah. The objective of this team was to | + | <p align="left" style="background-color:#ffffff;"> <br><br><br>Team TSH consisted of Emily, Helen and Sarah. The objective of this team was to express the thyroid stimulating hormone antagonist in two different forms. |
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+ | </br><br>These were with 'His tags' and without. Both of these constructs can be found on the <a href="https://2017.igem.org/Team:Cardiff_Wales/basicparts"> basic parts page</a>. This team was responsible for creating the <a href="https://2017.igem.org/Team:Cardiff_Wales/compositeparts"> level one constructs</a> with the 35S or LexA promoter containing the TSH or TSHH CDS. </br><br>The <a href="https://2017.igem.org/Team:Cardiff_Wales/basicparts"> level zero constructs</a> were put together using the type IIS restriction enzyme BsmB1 at 55 degrees C (Not 37!). These level 0 parts were then combined to create the level 1 constructs mentioned above using Bsa1 which (thankfully) does cut at 37 degrees. These constructs were used to transform <i> Agrobacterium </i>, which was infiltrated into our <i> Nicotiana benthamiana </i> plants. In adddition we worked with <a href="https://2017.igem.org/Team:Cardiff_Wales/Team_PlantP">Team Plant_P</a> to express TSHH using the plant promotors that they have generated as level 0 parts.</br><br>At the end of the project although we had not optimised the TSH protein expression, we know our tobacco transformations worked as demonstrated by the luciferase reporter gene work performed by <a href="https://2017.igem.org/Team:Cardiff_Wales/Team_Luc"> team luciferase</a> so we are hopeful that this will occur in the future. </br><br>Therefore we submited these <a href"https://2017.igem.org/Team:Cardiff_Wales/basicparts">TSH level 0 parts</a> to the registry after confirming them by sequencing! Finally although we ran out of time in the lab to optimse the expression assays, Sarah will be continuing this work in a future research project. | ||
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Latest revision as of 18:33, 30 October 2017