Difference between revisions of "Part:BBa K302012:Experience"

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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.  
 
|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.  
 
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|'''Graph2''':
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|'''Figure 2''':
 
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|[[Image:newcastle_no induction.jpg|600px]]
 
|[[Image:newcastle_no induction.jpg|600px]]
 
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|Graph 2 shows the percentage of cells at different lengths(μm)uninduced
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|Figure 2 shows the percentage of cells at different lengths(μm)uninduced
 
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|'''See below''': ''Bacillus subtilis 168'' cells (left) and non-induced cells(right)
 
|'''See below''': ''Bacillus subtilis 168'' cells (left) and non-induced cells(right)
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|[[Image:Teamnewcastle_yneA168BS.jpg|300px]][[Image:Teamnewcastle_noindBS.jpg|300px]]   
 
|[[Image:Teamnewcastle_yneA168BS.jpg|300px]][[Image:Teamnewcastle_noindBS.jpg|300px]]   
 
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|'''Graph3''':
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|'''Figure 3''':
 
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|[[Image:newcastle_0.2 induction.jpg|600px]]
 
|[[Image:newcastle_0.2 induction.jpg|600px]]
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|[[Image:Teamnewcastle_yneA168BS.jpg|300px]][[Image:Teamnewcastle_0.2indBS.jpg|300px]]
 
|[[Image:Teamnewcastle_yneA168BS.jpg|300px]][[Image:Teamnewcastle_0.2indBS.jpg|300px]]
 
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|'''Graph4''':
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|'''Figure 4''':
 
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|[[Image:newcastle_1IPTG.jpg|600px]]
 
|[[Image:newcastle_1IPTG.jpg|600px]]
 
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|Graph 4 shows the percentage of cells at different lengths(μm)induced at 1mM IPTG  
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|Figure 4 shows the percentage of cells at different lengths(μm)induced at 1mM IPTG  
 
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|'''See below''': ''Bacillus subtilis 168'' cells (left) and cells induced at 1mM IPTG(right)
 
|'''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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'''Figures 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. '''
  
 
===User Reviews===
 
===User Reviews===

Revision as of 16:04, 27 October 2010

This experience page is provided so that any user may enter their experience using this part.
Please enter 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).

Table1:

Stats: 168 yneA pMutin4 0μM IPTG pMutin4 1μM IPTG
Average: 1.34μm 3.53μm 1.74μm 3.19μm
Max: 2.30μm 6.00μm 3.62μm 9.77μm
Min: 0.55μm 1.31μm 0.88μm 1.14μm
Median: 1.33μm 3.27μm 1.62μm 2.66μm
Standard Deviation: 0.32μm 1.01μm 0.80μm 1.56μm
Figure1:
Distribution of cell lengths is not normal, so the mean is misleading; we are reporting the median instead.
Teamnewcastle yneA168.png
Teamnewcastle yneA168BS.jpgTeamnewcastle yneA1.jpgTeamnewcastle yneA.jpg
Figure1: shows statistics for populations of cells
  • overexpression of the yneA construct (ΔamyE:pSpac(hy)-oid::yneA(cells with YneA construct but no inhibitory regulation) ) leads to a longer cell length compared with our control Bacillus subtilis 168.
  • pMT4_0.0: YneA construct in pMutin4 vector with inhibition and no IPTG (ΔamyE:Pspac(hy)-oid::yneA::pMutin4)
  • pMT4_1.0: YneA construct in pMutin4 vector with inhibition and 1.0 μM IPTG (ΔamyE:Pspac(hy)-oid::yneA::pMutin4)
with inhibition cell lengths are comparable to Bacillus subtilis 168 at 0μM IPTG and longer with IPTG induction.
See Images below graph: Bacillus subtilis 168 cells (left),Bacillus subtilis expressing yneA(centre) and Bacillus subtilis overexpressing yneA(right)
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.
Figure 2:
Newcastle no induction.jpg
Figure 2 shows the percentage of cells at different lengths(μm)uninduced
See below: Bacillus subtilis 168 cells (left) and non-induced cells(right)
Teamnewcastle yneA168BS.jpgTeamnewcastle noindBS.jpg
Figure 3:
Newcastle 0.2 induction.jpg
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)
Teamnewcastle yneA168BS.jpgTeamnewcastle 0.2indBS.jpg
Figure 4:
Newcastle 1IPTG.jpg
Figure 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)
Teamnewcastle yneA168BS.jpgTeamnewcastle 1indBS2.jpg

Figures 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.

User Reviews

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