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Team:Biwako Nagahama/Project
From 2013.igem.org
Contents |
AgRePaper&E.coli-ink
Cellulose is used as raw material for paper, so our team experimented various ways to increase the amount of cellulose produced by agrobacterium and using it to make papers. For this we developed the different parts to insert into the system of agrobacterium. Among them are the genes used for expression of the curdlan. Similarly, genetic parts in order to increase the expression of the cellulose, along with the agrobacterium type binary vector were also developed . We are also working on recycling the produced paper by degrading the cellulose to D-Glucose using various enzymes. We worked for the preparation of the biological ink using the sperm whale's cells by genetically modification to increase amount of myoglobin. Then, we observed the change on the color of the product by altering the formation of myoglobin and the production amount of myoglobin with the insertion of T7 promoter to the cell system.
Agrepaper
PROBLEM PRESENTATION
①There are variety of issues that advanced in from today. For example Grobal warming,ozone layer destruction,biological extinction,an energy problem,desertification ,a crisis of water resources.There issues are coused by deforestation to make papers.
②How to solve it.
③Eenvironment issues can solve if not to use wood as well as burdon the environment. We need to know “What is the paper?” to think the method.
④The main component of paper is cellulose and hemicellulose.
⑤We were confirmed by Nakasima’s experiments that Agrobacterium make cellulose. Agrobacterium can’t make hemicellulose but cardlan Agrobacterium made has potential instead of hemicellulose, Ootaki perform confimatory experiment .As a result cardlan is able to instead of hemicellulose.
中島の実験
For this year’s project our team aimed at making the paper by using Agro bacterium. So, we confirmed whether carbohydrate developed from Agro bacterium could be used as the raw material for making paper.
Materials
Agro bacterium tumefaciens C58
Agro bacterium tumefaciens C58 bacterial liquid
＋LB medium 2 ml
＋ampicillin 2 µl
＋Agro bacterium tumefaciens C58
Congo red
LB medium
＋1.0％(w/v) Triptone 2 g
＋0.5％(w/v) yeast extract 1 g
＋1.0％(w/v) NaCl 2 g
Sample 1 curdlan 15 µl ＋Congo red 5 µl ＋Agro bacterium precipitationSample 1 curdlan 15 µl ＋Congo red 5 µl ＋Agro bacterium precipitationSample 1 curdlan 15 µl ＋Congo red 5 µl ＋Agro bacterium precipitation
Sample 2 de-ionized water 15 µl ＋Congo red 5 µl ＋Agro bacterium precipitation
We centrifuged Sample 1 and 2 in 5 minutes with cooled centrifuges(4 ℃,13000 rpm).
As a result, Sample 1 had red precipitates. Sample 2 rationally melted.
So, we confirmed the presence of carbohydrate in the precipitates obtained from Agro bacterium.
Next, we found that carbohydrate could be produced when soy milk is added to the fluid bacteria
So, we experimented for the confirmation of the above findings.
Sample 3 LB medium 7 ml ＋Agro bacterium＋soy 40 µl
Sample 4 LB medium 7 ml ＋Agro bacterium
Sample 5 LB medium 7 ml ＋soy 40 µl
Sample 6 LB medium 7 ml
We centrifuged Sample 1 and 2 in 5 minutes with cooled centrifuges(4 ℃,13000 rpm).
As a result, Sample 1 had red precipitates. Sample 2 rationally melted.
So, we confirmed the presence of carbohydrate in the precipitates obtained from Agro bacterium.
Sample 2 de-ionized water 15 µl ＋Congo red 5 µl ＋Agro bacterium precipitation
We centrifuged Sample 1 and 2 in 5 minutes with cooled centrifuges(4 ℃,13000 rpm).
As a result, Sample 1 had red precipitates. Sample 2 rationally melted.
So, we confirmed the presence of carbohydrate in the precipitates obtained from Agro bacterium.
Next, we found that carbohydrate could be produced when soy milk is added to the fluid bacteria<p/> <p>So, we experimented for the confirmation of the above findings.
Sample 3 LB medium 7 ml ＋Agro bacterium＋soy 40 µl
Sample 4 LB medium 7 ml ＋Agro bacterium
Sample 5 LB medium 7 ml ＋soy 40 µl
Sample 6 LB medium 7 ml
We cultivated Sample 3 ~ 6 at 28 ℃.for 12 hours
Then, Sample1 and 2 produced a white film like substance but Sample 3 and 4 did not.
We put the white films from Sample 1 and 2 into micro-tubes.
