Difference between revisions of "Team:Manchester/Results"

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Lorem ipsum dolor sit amet, consectetur adipiscing elit. Sed condimentum turpis ut aliquam pellentesque. Quisque consectetur quam lorem, ut varius elit mollis euismod. Fusce gravida orci metus, nec cursus dui pulvinar quis. Vivamus accumsan magna tortor, quis egestas magna faucibus in. Morbi enim sem, sollicitudin viverra viverra et, tempor sit amet justo. Quisque malesuada magna eu lorem egestas, quis molestie lectus sollicitudin. Donec varius magna vel eros faucibus, vel cursus eros condimentum. Donec justo lacus, tempus sed vestibulum quis, semper vitae nulla. Nam vel accumsan mi. Cras molestie, risus in hendrerit laoreet, sapien mauris ultricies enim, in fermentum ligula nibh a tellus. In vitae sollicitudin nibh. Praesent convallis consectetur vulputate. Vestibulum faucibus augue a pretium interdum. Phasellus eros felis, tincidunt eget odio a, imperdiet consequat ipsum. Duis bibendum mattis efficitur. Etiam auctor id elit at fringilla. Donec pretium, ex id pellentesque varius, risus lacus pellentesque tortor, non tincidunt eros libero vel mi. Morbi viverra tempor mattis. Curabitur mattis sed dolor in aliquet. Maecenas nec metus at nibh gravida commodo id vel dolor. Morbi commodo cursus lacus, et ullamcorper nibh lacinia in. Mauris lectus enim, sodales vel dolor eget, volutpat volutpat purus. Quisque vitae odio pellentesque, pulvinar lorem eu, congue ligula. Morbi magna sem, pharetra sed sagittis quis, aliquam sit amet purus. Aliquam eget est ut ligula facilisis gravida ut ut ante. Nullam quis venenatis nulla. Nam facilisis ex in ligula consequat posuere. Pellentesque id arcu mattis, gravida elit a, hendrerit ante. Integer mollis justo sed vestibulum vehicula.

So we designed all of the tag-promoter constructs with mCherry attached so that they could be visualized via fluorescent microscopy. We visualized the tag-promoter constructs using mCherry to check the distribution of the tag throughout the cell and to see the level of expression based on fluorescence.

As expected, without the expression of any Eut subunits, the tag showed a homogeneous distribution throughout the cells with no localization. The difference in fluorescence was most pronounced between low promoter and the other two as shown (in another figure? There’s a graph showing medium and high expression levels as very similar).

As medium promoter and high promoter had a similar level of expression, only low and high were combined with Eut subunits for visualization.

Levels of fluorescence from LowS were too low to properly visualize. HighS showed slightly heterogeneous distribution of fluorescence, the fluorescence was slightly granular with some brighter areas and some darker areas but no well-defined localization.

LowSMN images were very dim but visualization was possible. The fluorescence was granular but mainly heterogeneous throughout the cell. HighSMN showed quite well defined localization in a number of cells.

SMNLK was visualized with the low promoter as successful construction of HighSMNLK was not possible before we ran out of time. Fluorescence from LowSMNLK showed quite well-defined localization with a number of cells showing relatively round accumulations of fluorescent tag, suggesting proper BMC formation.

It is important to note that the majority of the accumulations seen occur at or near the end of the cells. This could be indicative of protein aggregates and not proper BMCs.