Difference between revisions of "Team:Macquarie Australia/Improve"

Line 12: Line 12:
 
<html>
 
<html>
 
<p align="justify">
 
<p align="justify">
−
We have improved the characterisation of the previously submitted part <a href="https://parts.igem.org/Part:BBa_K1998011”> BBa_K1998011 </a>. This was submitted by the Macquarie University iGEM team of 2016 and contains the <i>Chlamydomonas reinhardtii</i> Ferredoxin protein and Ferredoxin NADP+ reductase.
+
We have improved the characterisation of the previously submitted part <a href="http://parts.igem.org/Part:BBa_K1998011”> BBa_K1998011 </a>. This was submitted by the Macquarie University iGEM team of 2016 and contains the <i>Chlamydomonas reinhardtii</i> Ferredoxin protein and Ferredoxin NADP+ reductase.
 
<br>
 
<br>
 
<br>
 
<br>

Revision as of 04:44, 29 October 2017



menubanner

We have improved the characterisation of the previously submitted part
The expression of our proteins was then further confirmed by conducting an SDS-PAGE gel on the collected fractions with observed colour as can be seen in Figure 2.

Following this we conducted an enzymatic assay on the collected fractions to observe the activity of FNR using cytochrome C. The assay was performed using a master mix of 5 µM Ferredoxin, 250 µM cytochrome C and 0.2 µM NADPH. 5 µL of each fraction was added to the master mix and the change in absorbance was observed using a spectrometer at 550 nm. The observed change was of the FNR enzyme oxidising the NADPH to NADP+ while reducing the Ferredoxin. The electrons from the NADPH was then transported to the cytochrome C protein by the Ferredoxin protein. The reduction of cytochrome C is responsible for the observed change in absorbance. This can be seen in Figure 3 which shows activity of each fraction on the observed peaks from the anionic exchange chromatography.

Our results showed that the fractions B10 and B6 were the most active in reduction of cytochrome C.
We were able to not only express and purify the Ferredoxin and FNR proteins, but we also were able to prove that the proteins were capable of the oxidation of NADPH and reduction with the use of Ferredoxin.


GOLD SPONSORS


BRONZE SPONSORS


LOCATION


Faculty of Science and Engineering,
Macquarie University
Balaclava Road, North Ryde, NSW, 2109, Australia
E7B 350

CONTACT US

Email:
macquarie.australia@gmail.com

FOLLOW US ON