Team:UESTC-China/yanTeam

Feb 14
iGEM 2014 registration opens.
Feb 20
Site: West 304 of main building in UESTC
Team of iGEM 2014 in UESTC is founded.
Feb 21
Site: West 304 of main building in UESTC
We name our team Green Life;
Discuss the direction we will focus on for the iGEM 2014.
Feb 24
Site: West 304 of main building in UESTC
We meet the supervisors Yong Zhang, Xuelian Zheng, Lixia Tang, discussing the direction and acquiring their opinions of our project.

March 3
Site: West 304 of main building in UESTC
We firstly choose plant as our receptor, each member of our team give advices for the details we plan to do, such as which plant to use, tobacco, Arabidopsis or any other plant?
March 10
Site: West 304 of main building in UESTC
We talk with our supervisors and decide to use tobacco as our receptor for its simpleness as model organism.
March 17
Site: West 304 of main building in UESTC
We discuss the project and firstly decide to create a plant which can absorb formaldehyde largely by synthetic biology. And we choose Yasong Cui as our team leader.
March 24
Site: West 304 of main building in UESTC
Team leader assign task for us;
The first phase for our task begins, searching references and designing our logo.
March 31
We registered our team with the name of Green Life;
iGEM 2014 registration closes;
Team registration fee due.

April 9
Site: West 304 of main building in UESTC
Group meeting for the references and information we have researched last few days. Team leader Yasong Cui and Jingyao Lireport the key enzymes through the formaldehyde metabolic pathways. HPS, PHI are critical enzymes of RuMP pathway in formaldehyde assimilation. The references show that the transformation method is chloroplast transformation. The crucial enzyme in amplifying stomas of foliage.
April 13
Site: West 304 of main building in UESTC
Group meeting for the reference reading.
Mingyuan Wang report one gene (AdCP) of pollen sterility;
Jiao He report FDH which can amplify metabolic pathway in plant;
Yasong Cui report gene AHA2 which can amplify stoma of foliage.
April 16
Site: West 304 of main building in UESTC
Group meeting for the wiki constructing;
The responsible person of wiki, Zhiqiang Yan assign task for the rest.
April 20
Site: West 304 of main building in UESTC
Jie Li report Faldh which can amplify metabolic pathway in plant;
Rui Liu report some information about HPS, PHI;
Yasong Cui report some information about AHA2.
April 23
Site: West 304 of main building in UESTC
Vector construction and confirmation of the gene (HPS, PHI, FDH, Faldh, AHA2, AdCP) being used in our project.
April 23
Site: online
We exchange the team name Green Life into UESTC-China.
April 30
Site: The seventh middle school in Chengdu.
We introduce the concept and racing type of iGEM to the students in the seventh middle school who were the best students in Sichuan province.
April 30
Site: West 304 of main building in UESTC
We talk about the PrbcS-3C promoter sequence and the gene sequence of transit peptide of chloroplast.

DNA Distribution Kit sent to teams
May 01
iGEM 2014 Late registration closes
May 3
Site: West 304 of main building in UESTC
Decisionon the backbone vector.
The backbone named pXZY006 was proffered by one of our advisor Zhengyang Xie. And we designate it piGEM001.
Weixin Liu finds some chloroplast and cytoplasm transit peptide from Uniprot database. May 7
Site: West 304 of main building in UESTC
Take a group meeting and design the vectors we plan to use.
May 14
Site: West 304 of main building in UESTC
Group meeting.
We discussed some details in the designing of vector constructing;
At the end of the meeting, we started our experiments.
May 19
Site: West 121 of main building in UESTC
We extract pXZY006 plasmid from bacterium culture using TIANGEN kit.
Using SpeⅠand HpaⅠto do the double digestion of pXZY006 and F01 (contains HPS ) and do the ligation using T4 DNA ligase.
May 22
Site: West 121 of main building in UESTC
Do the transformation in competent recipient E.coli cell.
May 23
Site: West 121 of main building in UESTC
Repeat the experiment we do in May 19 and May 22 for the outcome is not that satisfactory.
Also we do not use the T4 ligase to do the ligation but by Gibson assembly.
May 28
Site: West 121 of main building in UESTC
Do the colony PCR to testify if the F01 has ligated to pXZY006.
And it comes out with positive clones, so the vector piGEM001 has been successfully constructed.
May 29
Site: West 304 of main building in UESTC
Group meeting;
Decide to build the second vector piGEM002.
Ligate F05, F02, and F03 to piGEM001 to construct piGEM002;
Ligate F06, F07 to piGEM002 to construct piGEM011.
May 30
Site: West 121 of main building in UESTC
Ligate F06 and F07 to iGEM001.
May 31
Site: West 121 of main building in UESTC
Do the transformation using heat shock.
The outcome is satisfactory that there is no clone in control group and thirteen clones in experimental group.

June 5
Site: West 121 of main building in UESTC
Do the colony PCR to testify if the F06 and F07 have ligated to piGEM001.
This ligation fail due to the outcome of colony PCR.
June 6
Site: West 121 of main building in UESTC
Repeating the experiment we did on May 31.
June 7
Site: West 121 of main building in UESTC
Do the transformation and colony PCR.
The outcome of colony PCR still demonstrate we fail again.
June 8-13
Site: West 121 of main building in UESTC
We do the ligation and transformation repeatedly, asking help from our advisors, finally we got the right outcome of piGEM011 (not the originally piGEM011 we planned to construct, used to be F06+F07+piGEM002 and now is F06+F07+piGEM001)
June 14
Site: West 121 of main building in UESTC
We ligate F02, F03, and F05 but fair.
June 15
Site: West 121 of main building in UESTC
We ligate F02, F03, and F05 but fair.
June 16
Site: The area before Sun Shine dining hall in UESTC
We made a knowledge quiz which contains a few questions concerning iGEM or synthetic biology to expand people's horizon.
June 19
Open Office Hours -- ask safety questions, meet other iGEMers!
June 23
About our Lab form due
June 17-30
Site: West 121 of main building in UESTC
We do the multiple ligation of F02, F03, and F05 repeatedly for changing the parameters in experiments but fair.

Vector construction

On August 6th, after about three months' hard work,we finish the section of vector construction. According to the plan,we are going to construct 11 gene expression vectors,including 2 backbone vectors, 6 single gene expression vectors and 3 multi-gene expression vectors. The main reason why it takes so long is that the gene AHA2 we synthetize from the company can't stay stable in our competent cell. Several methods are tried, but they don' work. So we have to give the gene up.

Visited by the schoolmaster

During the summer vacation, besides communicating with the students in SCU, there is another activity which worth a notice, our principal came to our laboratory and gave us a speech, which totally stimulated us to work harder and excited. The principal told us do not be scared to have the new idea, ‘cause the study we received today is to make us to speak out a new idea no matter it is wizard or not.

Questionaire

In order to popular iGEM in our country and do some research about our project, we made a questionnaire. We disseminated the questionnaire in many different places to make different range of people to participate this research, however, the students is still the most part of the engaged people.
In the questionnaire, we got three part needing people to sign in. through these three parts’ questions, we can find the viewpoints people showed to iGEM and our project.

Formaldehyde detection

On September 1st, when the eighth leaves of the first batch of tobacco seedings appear,we start the formal experiments of formaldehyde detection. Before that,some pre-tests are made based on some corresponding references. Finding the right amount of formaldehyde is the hardest part. By means of a series of experiments,we determine the final protocol which includes the qualitative detection and quantitative detection. From the experimental results,we can draw the conclusion that our super plants have remarkable formaldehyde tolerance and can dramatically reduce the concentration of formaldehyde in the air.

Products display

In the beginning of this new semester, our school held a science and culture activity. We caught this opportunity to advertise iGEM and our ultimate products, the plants which can absorb formaldehyde. The leaders and many teachers in our school also participated this activity. We displayed the plants which already owned the several genes which worked in the absorption of formaldehyde, to the people who visited our project. And the people visited our plant all said they want to owe one if there is already market available.