Difference between revisions of "Part:BBa K2623007:Experience"

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====Identification====
 
====Identification====
 
We performed a small amount of protein expression by SDS-PAGE.<br>
 
We performed a small amount of protein expression by SDS-PAGE.<br>
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<table><tr><th>[[Image:SAHS pro Gel 1.png|thumb|800px|Fig.1 The marker is on the lest, followed by our control group ( the BL21 with the empty plasmid), and the third well is the concentrated supernatant.]]</th><th></table><br>
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<table><tr><th>[[Image:SAHS pro Gel 1.png|thumb|800px|Fig.1 The marker is on the lest, followed by our control group ( the BL21 with the empty plasmid), and the third well is the concentrated supernatant. The unit of the marker is "Kd". ]]</th><th></table><br>
  
 
SAHS is an exogenous protein with a signal peptide in front of it that allows the protein to be secreted outside the membrane. But our site organism is E. coli, because there is a cell wall, so the protein can not be secreted out of the membrane. Therefore, we can hardly see our target protein in the supernatant. Therefore, we can only obtain the protein we need from the cell pellet. After that, we connected our SAHS protein to the pET-28a plasmid, induced expression, and obtained a large amount of our target protein by ultrasonic disruption of the bacterial cell pellet.<br>
 
SAHS is an exogenous protein with a signal peptide in front of it that allows the protein to be secreted outside the membrane. But our site organism is E. coli, because there is a cell wall, so the protein can not be secreted out of the membrane. Therefore, we can hardly see our target protein in the supernatant. Therefore, we can only obtain the protein we need from the cell pellet. After that, we connected our SAHS protein to the pET-28a plasmid, induced expression, and obtained a large amount of our target protein by ultrasonic disruption of the bacterial cell pellet.<br>

Latest revision as of 19:40, 17 October 2018

Applications of BBa_K2623007

Identification

We performed a small amount of protein expression by SDS-PAGE.

Fig.1 The marker is on the lest, followed by our control group ( the BL21 with the empty plasmid), and the third well is the concentrated supernatant. The unit of the marker is "Kd".

SAHS is an exogenous protein with a signal peptide in front of it that allows the protein to be secreted outside the membrane. But our site organism is E. coli, because there is a cell wall, so the protein can not be secreted out of the membrane. Therefore, we can hardly see our target protein in the supernatant. Therefore, we can only obtain the protein we need from the cell pellet. After that, we connected our SAHS protein to the pET-28a plasmid, induced expression, and obtained a large amount of our target protein by ultrasonic disruption of the bacterial cell pellet.
More information about our project can be found on our results page http://2018.igem.org/Team:XMU-China/Results

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