Difference between revisions of "Part:BBa K2623015:Experience"
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In our circuit of the build process, we have been doing nucleic acid gel electrophoresis to verify. After the loop is complete, sequencing verification. | In our circuit of the build process, we have been doing nucleic acid gel electrophoresis to verify. After the loop is complete, sequencing verification. | ||
https://static.igem.org/mediawiki/parts/1/1c/SAHS_1_Fig1.png | https://static.igem.org/mediawiki/parts/1/1c/SAHS_1_Fig1.png | ||
| + | https://static.igem.org/mediawiki/parts/2/28/SAHS_1_Fig2.png | ||
| + | After the fluorescence identification, we performed a small amount of protein expression by SDS-PAGE. | ||
| + | |||
| + | https://static.igem.org/mediawiki/parts/c/c5/SAHS_1_Fig3.png | ||
| + | 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. | ||
===User Reviews=== | ===User Reviews=== | ||
Revision as of 13:37, 29 September 2018
This experience page is provided so that any user may enter their experience using this part.
Please enter
how you used this part and how it worked out.
Applications of BBa_K2623015
In our circuit of the build process, we have been doing nucleic acid gel electrophoresis to verify. After the loop is complete, sequencing verification.
After the fluorescence identification, we performed a small amount of protein expression by SDS-PAGE.
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.
User Reviews
UNIQ8cdf695f760471e7-partinfo-00000000-QINU UNIQ8cdf695f760471e7-partinfo-00000001-QINU