Difference between revisions of "Part:BBa K3498029:Experience"

(Applications of BBa_K3498029)
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===Applications of BBa_K3498029===
 
===Applications of BBa_K3498029===
PvanCC promotor + Rsn-2 RBS + Rsn-2 + Terminator
 
 
This composite part is regulated by vanillic acid sensitive promoter PvanCC. This part contains an histidine tag used for purification. Al. assembled in pSB3K3 plasmid, a low number of copies plasmid
 
 
Introduction
 
 
Ranaspumin-2
 
Ranaspumin-2, also known as Rsn-2 is a recombinantly produced protein in E. coli Top 10. Its foamability capacity is being tested for fire fighting foam production purposes
 
 
Results.
 
 
Protein expression in E. coli Top 10
 
A characterization of the expression in E. coli was made. For this different concentrations of inducer (vanillic acid) and different final growing times were tested. Concentrations were determined by imageJ pixel density analyzer program
 
 
 
 
Figure 1: Induction conditions for Rsn-2
 
The results indicate that the best induction condition for Rsn-2 are 800 mM and 20 hours of growing at 37°C with a production of 0.4253 mg/mL
 
 
Foamability
 
 
The foamability of the expressed Rsn-2 was determined by testing the sonicated cultures, manually agitating a 15 mL corning tube containing 5 mL of the sonicated protein and observing the volume of the foam produced
 
Measurements were taken at 0, 1, 3, 5, 10, 15 and 30 minutes, after that, the measurements were taken every 30 minutes until 12 hours after the agitation
 
 
Figure 2: Water (negative control), Rsn-2, Rsn-3, Rsn 3-5, SDS (positive control), all agitated 30 second
 
 
The foamability is obtained by FV/SV where FV is foam volume and SV is solution volume. Ranaspumin-2 made a Foam volume of 4 mL with a 5 mL solution, hence its foamability is of 0.8 (relative number)
 
 
 
Figure 3: Foamability of Rsn-2
 
 
Passing the 12 hours the behaviour of the foam was analyzed in order to see how durable it is
 
 
 
Figure 4: Foam volume over 12 hours
 
 
It took 8.5 hours for the foam to go from 4 mL to 2 mL (Its half life), and at the final measurement it still had 1 mL of foam
 
  
 
===User Reviews===
 
===User Reviews===

Revision as of 15:14, 20 October 2021


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Applications of BBa_K3498029

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