Difference between revisions of "Team:SBS SH 112144/Composite Part"
Yinchizhou18 (Talk | contribs) |
Yinchizhou18 (Talk | contribs) |
||
| Line 4: | Line 4: | ||
<h1>Composite Part</h1> | <h1>Composite Part</h1> | ||
<h4> Composite Tagged Lysozyme Gene</h4> | <h4> Composite Tagged Lysozyme Gene</h4> | ||
| − | <img src="https://static.igem.org/mediawiki/2018/4/4e/T--SBS_SH_112144--fused_tagged_lysozyme.png" width= | + | <img src="https://static.igem.org/mediawiki/2018/4/4e/T--SBS_SH_112144--fused_tagged_lysozyme.png" width=950 height=400> |
<p>Description: Our composite part within prefix and suffix consists of the most commonly used promoter, RBS, 6x His tag and terminator, as well as our functional coding sequence --- cyanophage lysozyme gene. This composite part is assembled through Gibson Assembly; then, it is fused with linearized vector, pSB1C3 backbone, via infusion. Ultimately, we successfully extract plasmids which contain our target gene from transformed bacteria. Furthermore, we purposely add 6x histag into our sequence in order to improve the efficacy and efficiency of expression and purification; it will also strengthen effects of binding to Ni-NTA column, making it possible to immobilize proteins and lyse cyanobacteria repeatedly in our prototype device. </p> | <p>Description: Our composite part within prefix and suffix consists of the most commonly used promoter, RBS, 6x His tag and terminator, as well as our functional coding sequence --- cyanophage lysozyme gene. This composite part is assembled through Gibson Assembly; then, it is fused with linearized vector, pSB1C3 backbone, via infusion. Ultimately, we successfully extract plasmids which contain our target gene from transformed bacteria. Furthermore, we purposely add 6x histag into our sequence in order to improve the efficacy and efficiency of expression and purification; it will also strengthen effects of binding to Ni-NTA column, making it possible to immobilize proteins and lyse cyanobacteria repeatedly in our prototype device. </p> | ||
<p>Length: 773 bp</p> | <p>Length: 773 bp</p> | ||
| Line 12: | Line 12: | ||
<p>Description: The second part is utilized to demonstrate the pure effects of lysozyme in our function test--- cyanobacterial lysis reaction. Mainly applied in function test, this composite part without 6x his tag was fused with pSB1C3 backbone and then was successfully expressed in E.coli. The lysis reaction was then conducted in the solution with both cultivated cyanobacteria and expressed proteins. In this way, we could examine and quantify the lysis results, confirming the efficacy of cyanophage lysozyme. </p> | <p>Description: The second part is utilized to demonstrate the pure effects of lysozyme in our function test--- cyanobacterial lysis reaction. Mainly applied in function test, this composite part without 6x his tag was fused with pSB1C3 backbone and then was successfully expressed in E.coli. The lysis reaction was then conducted in the solution with both cultivated cyanobacteria and expressed proteins. In this way, we could examine and quantify the lysis results, confirming the efficacy of cyanophage lysozyme. </p> | ||
<p>Length: 747 bp</p> | <p>Length: 747 bp</p> | ||
| + | |||
| + | <p><a href='https://2018.igem.org/Team:SBS_SH_112144/Experiments'>Our experience and application of composite parts</a ></p> | ||
| + | |||
</html> | </html> | ||
Revision as of 00:26, 16 October 2018
Composite Part
Composite Tagged Lysozyme Gene
Description: Our composite part within prefix and suffix consists of the most commonly used promoter, RBS, 6x His tag and terminator, as well as our functional coding sequence --- cyanophage lysozyme gene. This composite part is assembled through Gibson Assembly; then, it is fused with linearized vector, pSB1C3 backbone, via infusion. Ultimately, we successfully extract plasmids which contain our target gene from transformed bacteria. Furthermore, we purposely add 6x histag into our sequence in order to improve the efficacy and efficiency of expression and purification; it will also strengthen effects of binding to Ni-NTA column, making it possible to immobilize proteins and lyse cyanobacteria repeatedly in our prototype device.
Length: 773 bp
Composite Lysozyme Gene
Description: The second part is utilized to demonstrate the pure effects of lysozyme in our function test--- cyanobacterial lysis reaction. Mainly applied in function test, this composite part without 6x his tag was fused with pSB1C3 backbone and then was successfully expressed in E.coli. The lysis reaction was then conducted in the solution with both cultivated cyanobacteria and expressed proteins. In this way, we could examine and quantify the lysis results, confirming the efficacy of cyanophage lysozyme.
Length: 747 bp