Coding

Part:BBa_K1317003:Experience

Designed by: Mounir Benkoulouche   Group: iGEM14_Bordeaux   (2014-10-07)
Revision as of 18:14, 7 October 2014 by Mounir (Talk | contribs) (→‎Applications of BBa_K1317003)

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Applications of BBa_K1317003

The protein ELP has been produced in BL21 strain of E.coli. After phase cycling purification it is possible to spin the protein into a wire, thanks to the wet-spinning method. The ELP can have different length regarding the number of copy of the gene. In these experiments, we used 40mer ELP (assemble of two gene copies).

Polymer formation trial:

In order to obtain polymer wires, we carried out severeal tests by mixing the ELP with different solvants. The following results have been observed:

- Sodium Sulfate (Na2So4) at 320 g/L => cottony aspect of the fiber, the structure is not stable

- Diethyl ether (C2H5)2O => No reaction because the drop is trapped. This solvent is not miscible with the polymer

- Acetone (CH3COCH3) => Formation of a white ring

- Heated Sodium Chloride (NaCl 5M) => Formation of a white cloud due to a lack of cohesiveness

- Calcium Chloride (CaCl2 15%) => 1.25% alginate mixture with ELP40 at 10mg/mL: a white-colored fiber is obtained This last trial has been performed with 0.75% alginate which seems to be the best experimental condition to get fibers from the ELP

Phase transition trial:

After getting the ELP40 fiber, it is necessary to wash it with hot water for extractio, the fiber could indeed dissolve at low temperature.

- Control remains transparent

- The aspect of the fiber changes at ambient temperature (from white to transprent)

Thanks to the mixture alginate + ELP40 (10 mg/L), we could observe the formation of the polymer and it can be deduced that: - In presence of salt, product is losing cohesiveness

- With contact of hot water, the product is contracting

You can see the results on the phase transition on this video: https://www.youtube.com/watch?v=xoTQkoNoZu0

Wet-spinning and mechanical testing of fibers properties

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