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+ | {{NCKU_Tainan/header}} {{NCKU_Tainan/navbar}} {{NCKU_Tainan/style}} | ||
+ | <html> | ||
+ | |||
+ | <head> | ||
+ | <link rel="stylesheet" href="https://2018.igem.org/Template:NCKU_Tainan/css/improve?action=raw&ctype=text/css"> | ||
+ | </head> | ||
+ | |||
+ | <body> | ||
+ | |||
+ | <!--Page_Content--> | ||
+ | <div class="container content"> | ||
+ | <h1 class="head4">Improve</h1> | ||
+ | <div class="navbar-example"> | ||
+ | <div class="row"> | ||
+ | <div class="col-10"> | ||
+ | <div data-spy="scroll" data-target="#sidelist" data-offset="0" class="scrollspy-example"> | ||
+ | <div class="container"> | ||
+ | |||
+ | </br></br> | ||
+ | <h3>Part Improvement</h3> | ||
+ | <p class="pcontent"> | ||
+ | During the designation of the experiment for our pH alert system, we have concluded | ||
+ | that we should try to improve the expression of our pH-sensitive promoter, promoter | ||
+ | gadA. The motivation for us to do this is because our team think that if we can build a | ||
+ | system which could observe the situation in our device by looking at the color change | ||
+ | of the medium, it should be an economic friendly as there is no more extra charges to | ||
+ | send your sample to someway for checking and also wasting a lot of time for waiting the | ||
+ | result. But according to the expression of promoter gadA from the previous team did, it | ||
+ | is not enough to be observed by using our eyes. | ||
+ | </p></br> | ||
+ | |||
+ | <div id="pt"> | ||
+ | </br> | ||
+ | <p class="pcontent"> | ||
+ | Based on our research, RiboJ first described as it is an insulator in genetic | ||
+ | construct but there has no any data shown this insulation effected the downstream | ||
+ | genetic parts. In 2018, there is a team have proved that RiboJ could increase the | ||
+ | expression of downstream gene and there is the team member, Kalen P Clifton#1, | ||
+ | Ethan M Jones#1, Sudip Paudel1, John P Marken2, Callan E Monette1, Andrew D | ||
+ | Halleran2, Lidia Epp1, Margaret S Saha1*. Hence, our team, NCKU_Tainan have desided | ||
+ | to enhance the expression and specificity of promoter gadA by adding a gene, RiboJ | ||
+ | at the downstream of promoter gadA. | ||
+ | </p></br> | ||
+ | </div> | ||
+ | |||
+ | <div id="pt"> | ||
+ | </br> | ||
+ | <p class="pcontent">For the function determination, we have measured the fluorescent | ||
+ | density of the plasmid with and without RiboJ in different pH environment. We | ||
+ | incubated the bacteria in pH adjacent M9 medium and measure the fluorescent | ||
+ | intensity (absorbance: 480nm, excitation: 510 nm) in a short period of time.</p></br> | ||
+ | </div> | ||
+ | |||
+ | |||
+ | |||
+ | </br></br></br></br> | ||
+ | <h3>Experiment Result</h3> | ||
+ | <div id="pt"> | ||
+ | <p class="pcontent">This is the third year since the establishment of iGEM | ||
+ | NCKU-Tainan team. We have raised the awareness of synthetic biology in our | ||
+ | campus. Mrs. Jenny Su and the College of Engineering decided to give us a | ||
+ | headquarter which contains a meeting room and a lab. To meet the lab standard | ||
+ | of CDC (Center for Disease Control), we made contact with Mrs. Liu Yee Fen, the | ||
+ | contact person of Center for Occupational safety and Health and Environmental | ||
+ | Protection, which is the authorized unit in our campus responsible for lab | ||
+ | safety and security. With the assistance of our instructor Dr. Ng, we | ||
+ | successfully certify our lab to BSL1 saftey level one lab. Mrs. Liu gave us | ||
+ | some lab security instructions and advice. She also suggested that each of our | ||
+ | members should attend the biosafety lab training program. | ||
+ | </p> | ||
+ | </div> | ||
+ | |||
+ | <div id="pt"> | ||
+ | <p class="pcontent">Expect the lab in iGEM headquarter, wet team has constructed | ||
+ | out design in various labs whose supervisors are Dr. I-Son Ng, Dr. I-Hsiu | ||
+ | Huang, Dr. Han-Ching Wang, and Dr. Hashimoto Masayuki. The labs mentioned above | ||
+ | are all certified biosafety level one.</p></br> | ||
+ | </div> | ||
+ | |||
+ | |||
+ | |||
+ | </br></br></br></br> | ||
+ | <h3>Experiment Result</h3> | ||
+ | |||
+ | <div class="row"> | ||
+ | <div class="col-md-6 col-md-push-6"> | ||
+ | <div class="col-md-6 col-md-pull-6 appdiv"> | ||
+ | <img class="appimg" src="https://static.igem.org/mediawiki/2018/2/2f/T--NCKU_Tainan--home_42844661_1916518971978222_1636077502608703488_n.png"> | ||
+ | </div> | ||
+ | <p class="pcontent" style="margin-top: 200px"> | ||
+ | Figure 1: The fluorescent intensity expressed by PgadA of 24 hours in four | ||
+ | different pH value of M9 medium. | ||
+ | </p> | ||
+ | </div> | ||
+ | </div> | ||
+ | |||
+ | <div class="row"> | ||
+ | <div class="col-md-6 col-md-push-6"> | ||
+ | <div class="col-md-6 col-md-pull-6 appdiv"> | ||
+ | <img class="appimg" src="https://static.igem.org/mediawiki/2018/4/41/T--NCKU_Tainan--home_42887276_674354272947210_4995870134384984064_n.png"> | ||
+ | </div> | ||
+ | <p class="pcontent" style="margin-top: 200px"> | ||
+ | Figure 2: The fluorescent intensity expressed by PgadA-RiboJ of 24 hours in | ||
+ | four different pH value of M9 medium. | ||
+ | </p> | ||
+ | </div> | ||
+ | </div> | ||
+ | |||
+ | <img class="contentimg" src="https://static.igem.org/mediawiki/2018/9/9f/T--NCKU_Tainan--home_42912088_2204785603177136_6518878486872981504_n.png"> | ||
+ | <p class="pcontent">Figure 3: The fluorescent intensity between PgadA and PgadA-RiboJ in pH | ||
+ | 6 and pH 7.</p> | ||
+ | |||
+ | </br></br></br></br> | ||
+ | <h3>Experiment Result</h3> | ||
+ | <div id="pt"> | ||
+ | </br> | ||
+ | <p class="pcontent"> | ||
+ | Based on our functional determination test results, the improvement has succeeded | ||
+ | as RiboJ really could enhance the expression and specificity of downstream gene. | ||
+ | </p></br> | ||
+ | </div> | ||
+ | |||
+ | |||
+ | </br></br></br></br></br></br> | ||
+ | |||
+ | </div> | ||
+ | |||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | </body> | ||
+ | |||
+ | </html> | ||
+ | {{NCKU_Tainan/footer}} |
Revision as of 13:19, 2 October 2018