Difference between revisions of "Part:BBa K3498010:Experience"

Latest revision as of 21:36, 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_K3498010

User Reviews

UNIQ7aa3ed0ba8282a90-partinfo-00000000-QINU UNIQ7aa3ed0ba8282a90-partinfo-00000001-QINU

•••••

IGEM FCB-UANL

PvanCC promotor + Rsn-3 RBS + Rsn-3.

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. This part does not contain terminator

Introduction

Ranaspumin-3.

Ranaspumin-3, also known as Rsn-3 is recombinantly produced protein in E. coli Top 10. Its foam stability 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

First, different concentrations were tested for a short period of time, resulting thath 1000 mM was the best concentration for production, afterwards, this concnetration was tested for different hours

Figure 1: Induction conditions for Rsn-3

The results indicate that the best induction condition for Rsn-3 are 1000 mM and 12 hours of growing at 37°C with a production of 0.5070 mg/mL

Foamability

The foamability of the expressed Rsn-3 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-3 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-3

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

After 12 hours the final volume of Rsn-3 was of 2.5, it means that the half life of its foam is even longer

@@ Line 46: / Line 46: @@
-Figure 1: Induction conditions for Rsn-3
+[[File:T-FCB-UANL--RSN3-I.png|200px|thumb|left|Figure 1: Induction conditions for Rsn-3]]
 The results indicate that the best induction condition for Rsn-3 are 1000 mM and 12 hours of growing at 37°C with a production of 0.5070 mg/mL
@@ Line 56: / Line 82: @@
-Figure 2: Water (negative control), Rsn-2, Rsn-3, Rsn 3-5, SDS (positive control), all agitated 30 second
+[[File:T--FCB-UANL--PE.png|200px|thumb|left|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 mL with a 5 mL solution, hence its foamability is of 0.8 (relative number)
-Figure 3: Foamability of Rsn-3
+[[File:T--FCB-UANL--E-GC.png|200px|thumb|left|Figure 3: Foamability of Rsn-3]]
 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
+[[File:T--FCB-UANL-E-GB.png|200px|thumb|left|Figure 4: Foam volume over 12 hours]]
 After 12 hours the final volume of Rsn-3 was of 2.5, it means that the half life of its foam is even longer