Difference between revisions of "Part:BBa K3033018"

Revision as of 02:12, 22 October 2019

OPHT Capture system version 1: 2-Tyrosine capturing domain

Overview

Our first and main feature of the whole Self-Activating Nanoparticle E.coli organism (SANCE). The Surface Nanoparticle Capturing Protein (SNCP). This variant name of this SNCP is OPHT Capture system version 1: 2-Tyrosine capturing domain The general overview of the whole feature is shown in Figure 1 below.

Figure 1: Overview of the mechanism and where our SNCP translocates in our bacteria

Our surface nanoparticle capturing protein (SNCP) has a couple of components as shown in figure 2. An anchoring protein. A linker enzyme. Next is the presence of 6-histidine tag. Finally, the capturing domain. Our inspiration for this capturing system is from the Mefp-5 protein that is found on the foot of the mussels. This inspiration protein has L-Dopa and histidine repeats in them thus we want to express L-Dopa as our capturing domain as it has a sticky property.

Figure 1: Overview of the mechanism and where our SNCP translocates in our bacteria

Our system would primarily be anchored onto the cell membrane of the bacteria using the outer membrane protein called OmpW derived from E. coli itself, it forms an 8-stranded beta-barrel with a long and narrow hydrophobic channel. We chose this as our anchoring protein as it is very stable and is able to carry our other protein onto the surface of the bacteria. We have truncated this version of OmpW at the 191st amino acid as it has been shown to be stably anchored onto the cell membrane while being able expose the rest of the proteins onto the outside of the cell membrane. The next component is the Perhydrolase which was derived from the organism Pseudomonas aeruginosa. According to our adviser, this would be enhancing our capturing system’s capability thorough the process of epoxidation. The next component is the 6- histidine tag which would be used to check the protein expression.

Protein Expression tests

Functional Tests

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 1511
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

@@ Line 4: / Line 4: @@
 ===Overview===
-[[File:T--UM_Macau--Feature_1.png|800px|thumb|centre|Figure 1: Overview of the mechanism and where our SNCP translocates in our bacteria
+Our first and main feature of the whole Self-Activating Nanoparticle E.coli organism (SANCE). The Surface Nanoparticle Capturing Protein (SNCP). This variant name of this SNCP is OPHT Capture system version 1: 2-Tyrosine capturing domain The general overview of the whole feature is shown in Figure 1 below.
+[[File:T--UM_Macau--Feature_1.png|500px|thumb|centre|Figure 1: Overview of the mechanism and where our SNCP translocates in our bacteria
 ]]
+Our surface nanoparticle capturing protein (SNCP) has a couple of components as shown in figure 2. An anchoring protein. A linker enzyme. Next is the presence of 6-histidine tag. Finally, the capturing domain. Our inspiration for this capturing system is from the Mefp-5 protein that is found on the foot of the mussels. This inspiration protein has L-Dopa and histidine repeats in them thus we want to express L-Dopa as our capturing domain as it has a sticky property.
+[[File:T--UM_Macau--Figure_2.png|500px|thumb|centre|Figure 1: Overview of the mechanism and where our SNCP translocates in our bacteria
+]]
+Our system would primarily be anchored onto the cell membrane of the bacteria using the outer membrane protein called OmpW derived from E. coli itself, it forms an 8-stranded beta-barrel with a long and narrow hydrophobic channel. We chose this as our anchoring protein as it is very stable and is able to carry our other protein onto the surface of the bacteria. We have truncated this version of OmpW at the 191st amino acid as it has been shown to be stably anchored onto the cell membrane while being able expose the rest of the proteins onto the outside of the cell membrane.
+The next component is the Perhydrolase which was derived from the organism Pseudomonas aeruginosa. According to our adviser, this would be enhancing our capturing system’s capability thorough the process of epoxidation. The next component is the 6- histidine tag which would be used to check the protein expression.