Difference between revisions of "Part:BBa K302012:Experience"
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|Distribution of cell lengths is not normal, so the mean is misleading; we are reporting the median instead. | |Distribution of cell lengths is not normal, so the mean is misleading; we are reporting the median instead. | ||
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|Graph 1 shows that overexpression (no Lac repression) of the ''yneA'' gene (Δ''amyE'':pSpac(hy)-oid::''yneA'') leads to a longer cell length compared with our control ''Bacillus subtilis 168''. | |Graph 1 shows that overexpression (no Lac repression) of the ''yneA'' gene (Δ''amyE'':pSpac(hy)-oid::''yneA'') leads to a longer cell length compared with our control ''Bacillus subtilis 168''. | ||
Revision as of 11:43, 27 October 2010
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how you used this part and how it worked out.
Applications of BBa_K302012
Characterisation by Team Newcastle 2010
We integrated our part into the Bacillus subtilis 168 chromosome at amyE (using the integration vector pGFP-rrnB) and selected for integration by testing for the ability to hydrolyse starch. Homologous recombination at amyE destroys endogenous expression of amylase. Colonies that are not able to break down starch on agar plate do not have a white halo when exposed to iodine.
The part was co-transcribed with gfp fluorescent marker by transcriptional fusion after the yneA coding sequence.
We characterised the part first without, and then with, LacI repression (using the integration vector pMutin4 to integrate lacI into the Bacillus subtilis 168 chromosome).
| Graph1: |
| Distribution of cell lengths is not normal, so the mean is misleading; we are reporting the median instead. |
|
| Graph 1 shows that overexpression (no Lac repression) of the yneA gene (ΔamyE:pSpac(hy)-oid::yneA) leads to a longer cell length compared with our control Bacillus subtilis 168. |
| See below: Bacillus subtilis 168 cells (left),Bacillus subtilis expressing yneA(centre) and Bacillus subtilis overexpressing yneA(right) |
  
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| The images we have taken this data from had very different numbers of cells, so the cells counts are misleading therefore we are reporting the proportions of cells at a given length. |
| Graph2: |
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| Graph 2 shows the percentage of cells at different lengths(μm)uninduced |
| See below: Bacillus subtilis 168 cells (left) and non-induced cells(right) |
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| Graph3: |
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| Graph 3 shows the percentage of cells at different lengths(μm)induced at 0.2mM IPTG |
| See below: Bacillus subtilis 168 cells (left) and cells induced at 0.2mM IPTG(right) |
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| Graph4: |
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| Graph 4 shows the percentage of cells at different lengths(μm)induced at 1mM IPTG |
| See below: Bacillus subtilis 168 cells (left) and cells induced at 1mM IPTG(right) |
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Graphs 2,3 and 4 show a greater proportion of cells at a higher concentration of IPTG(1mM IPTG), compared with Bacillus subtilis 168 our control population.
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