Difference between revisions of "Part:BBa K3498030:Experience"
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75 mM seemed to be the best candidate for the expression of ranaspumin 3-5, so it was tested out for 24 hours
 
75 mM seemed to be the best candidate for the expression of ranaspumin 3-5, so it was tested out for 24 hours
 +
 +
 +
https://parts.igem.org/File:T-FCB-UANL--RSN3-5-I.png
  
 
Figure 1: Induction conditions for Rsn-3-5
 
Figure 1: Induction conditions for Rsn-3-5
 +
  
 
The results indicate that the best induction condition for Rsn-3-5 are 75 mM and 12 hours of growing at 37°C with a production of 0.2014 mg/mL
 
The results indicate that the best induction condition for Rsn-3-5 are 75 mM and 12 hours of growing at 37°C with a production of 0.2014 mg/mL
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The foamability of the expressed Rsn-3-5 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  
 
The foamability of the expressed Rsn-3-5 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
 
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
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 +
 +
https://parts.igem.org/File:T--FCB-UANL--PE.png
  
 
Figure 2: Water (negative control), Rsn-2, Rsn-3, Rsn 3-5, SDS (positive control), all agitated 30 second
 
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-3 made a Foam volume of 4.5 mL with a 5 mL solution, hence its foamability is of 0.9 (relative number)
 
The foamability is obtained by FV/SV where FV is foam volume and SV is solution volume. Ranaspumin-3 made a Foam volume of 4.5 mL with a 5 mL solution, hence its foamability is of 0.9 (relative number)
 +
 +
 +
https://parts.igem.org/File:T--FCB-UANL--E-GC.png
  
 
Figure 3: Foamability of Rsn-3-5
 
Figure 3: Foamability of Rsn-3-5
 +
  
 
Passing the 12 hours the behaviour of the foam was analyzed in order to see how durable it is
 
Passing the 12 hours the behaviour of the foam was analyzed in order to see how durable it is
 +
 +
 +
https://parts.igem.org/File:T--FCB-UANL-E-GB.png
  
 
Figure 4: Foam volume over 12 hours
 
Figure 4: Foam volume over 12 hours
 +
  
 
It took 9 hours for the foam to go from 4.5 mL to  1.5 mL (lower than the half life), and at the final measurement it still had 1 mL of foam
 
It took 9 hours for the foam to go from 4.5 mL to  1.5 mL (lower than the half life), and at the final measurement it still had 1 mL of foam

Revision as of 20:12, 20 October 2021


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_K3498030

User Reviews

UNIQc0ceb7cba9ec3b21-partinfo-00000000-QINU UNIQc0ceb7cba9ec3b21-partinfo-00000001-QINU

•••••

IGEM FCB-UANL

PvanCC promotor + Rsn-3 RBS + Rsn-3 + Rsn-4 RBS + Rsn-4 + Rsn-5 RBS + Rsn-5 + Terminator.

This composite part is regulated by vanillic acid sensitive promoter PvanCC. This part contains an histidine tag used for purification. All assembled in pSB3K3 plasmid, a low number of copies plasmid

Introduction

Ranaspumin-3-5.

Ranaspumin-3-5, also known as Rsn-3-5 is recombinantly produced protein in E. coli Top 10. Its foam stability capacity is being tested for fire fighting foam production purposes. It is believed that the stability effect improves with the three ranaspumin working together

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

Firstly 100 and 1000 mM concentrations were tested, but with higher concentrations the expressed protein was lower, so lower concentrations were tested out

75 mM seemed to be the best candidate for the expression of ranaspumin 3-5, so it was tested out for 24 hours


https://parts.igem.org/File:T-FCB-UANL--RSN3-5-I.png

Figure 1: Induction conditions for Rsn-3-5


The results indicate that the best induction condition for Rsn-3-5 are 75 mM and 12 hours of growing at 37°C with a production of 0.2014 mg/mL

Foamability

The foamability of the expressed Rsn-3-5 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


https://parts.igem.org/File:T--FCB-UANL--PE.png

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-3 made a Foam volume of 4.5 mL with a 5 mL solution, hence its foamability is of 0.9 (relative number)


https://parts.igem.org/File:T--FCB-UANL--E-GC.png

Figure 3: Foamability of Rsn-3-5


Passing the 12 hours the behaviour of the foam was analyzed in order to see how durable it is


https://parts.igem.org/File:T--FCB-UANL-E-GB.png

Figure 4: Foam volume over 12 hours


It took 9 hours for the foam to go from 4.5 mL to 1.5 mL (lower than the half life), and at the final measurement it still had 1 mL of foam