Difference between revisions of "Part:BBa K2623015:Experience"

(→‎Identification)
(→‎Identification)
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After the fluorescence identification, we performed a small amount of protein expression by SDS-PAGE.<br>
 
After the fluorescence identification, we performed a small amount of protein expression by SDS-PAGE.<br>
 
<br>
 
<br>
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[[Image:SAHS_pro_Gel_1.png|thumb|400px|Fig.3]]<br>
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[[Image:SAHS_pro_Gel_1.png|thumb|800px|Fig.3]]<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>
 
More information about our project can be found on our results page.
 
More information about our project can be found on our results page.

Revision as of 08:56, 6 October 2018

Applications of BBa_K2623015

Identification

In our circuit of the build process, we have been doing nucleic acid gel electrophoresis to verify. After the loop is complete, sequencing verification.(Fig.1.2)

Fig.1
Fig.2


After the fluorescence identification, we performed a small amount of protein expression by SDS-PAGE.

Fig.3

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.

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