Current problems2012.igem.org/files/presentation/NYMU-Taipei_Championship.pdf · Achievements We...

Current problems

Population↑ Pollution↑

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The Solution: Conquering Venus &

COOL!!

NYMU-Taipei

But...

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1. Venus’ atmosphere…

2. Need energy, need food!!

CO2 SO2

NYMU-Taipei

Atmosphere Perform

Long-lasting

CO2

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H2S SO2

How about using H2S as energy source? → Sulfide-Quinone Reductase

Lots of CO2, but no O2 →Photosynthesis →Cyanobacteria

Becoming Venusians → Endosymbiosis, INV-LLO Bba_K299812 iGEM10_Warsaw (2010-10-13)

SO2 as a toxic gas →Sulfite reductase

Practical Application →Purifying polluted air

Overcoming difficulties

How about using H2S as energy source? → Sulfide-Quinone Reductase

NYMU-Taipei

Synechocystis PCC7942

Sulfite reductase

SO2

Sulfide quinone

reductase (sqr)

chloroplast +

S + (CH2O)n

C6H12O6

For Host Cell to

Use

Whole picture of our project: Inside of the Stem Cell

Glut

CO2

H2S

SO2

CO2

H2S

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Nucleus

Synechocystis PCC7942

How to become Venusians?

Desulfovibrio desulfuricans

Synechococcus sp. PCC 7002

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Engineered Synechocystis PCC7942

How to become Venusians?

Glut (BBa M11036 )

Invasin-listeriolysin O

(BBa_K299812)

Into the

Stem cell 7 NYMU-Taipei

CO2

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H2S SO2 SO2 as a toxic gas →Sulfite reductase

Sulfur Dioxide (SO2) Reducer

Solved!!

Synechocystis

PCC7942

NYMU-Taipei

Sulfur Dioxide (SO2) Reducer: The Concept

Desulfovibrio desulfuricans Pseudomonas aeruginosa

Dsr Sulfite reductase CysI Sulfite reductase

SO2 HSO3- H2S

dissolve in water Sulfite reductase

9 NYMU-Taipei

H2S serial dilution

and standard curve detection

When H2S exist,

DPDA solution turn

turbid.

H2S producing assay

Lavilla I,Pena-Pereira .F et al, Analytica Chimica Acta 647 (2009) 112–116

Sulfur Dioxide (SO2) Reducer: The Experiment

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WORK Sulfur Dioxide (SO2) Reducer: The Results

11 NYMU-Taipei In 500μ M K2SO4 medium

Experiment OK!! More H2S!!

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H2S SO2

Using H2S to generate energy How about using H2S as energy source? → Sulfide-Quinone Reductase

Solved!!

Synechocystis

PCC7942

Solved!!

Sulfite reductase

NYMU-Taipei

Sulfite reductase Sulfide quinone

reductase(sqr)

chloroplast

Synechocystis PCC7942

H2S

SO2

+

e-

H2S: Novel Energy Currency

NYMU-Taipei

S + (CH2O)n

CO2

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Sulfur Version of

Photosynthesis H2S S+2H++energy

SQR

NYMU-Taipei

Using H2S to generate energy: The Concept

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2H2O O2 +4H++energy

Cyanobacteria

DCMU challenged assay

NYMU-Taipei

Using H2S to generate energy: The Results

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sqr has

advantage

DCMU Cyanobacteria growth

Sodium Sulfide consumption in E. coli SQR grows better!

Flow cytometer – Exact cell number Auto-fluorescence assay method (2011

Schulze )

sqr strain has

larger cell number

and stronger

fluorescence signal

Plate Count

sqr strain

Wild type

sqr strain has more colonies

NYMU-Taipei

Using H2S to generate energy: The Results

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Sodium Sulfide

consumption in E. coli

Sodium Sulfide leftovers

after 48 hours

2.2 times!

sqr strain consumed

most of the sodium

sulfide WORK

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H2S SO2

Apoptosis

Creating Venusians

Becoming Venusians → Endosymbiosis

Solved!!

Synechocystis

PCC7942

Solved!!

Sulfite reductase

Solved!! sqr

NYMU-Taipei

Invasin(inv) from Yersinia pestis

HEp-2 cell

Inv+

E.coli

listeriolysin O(llo) from

Listeria monocytogenes

HeLa cell

llo+ E.coli

[1]Isberg, R.R., and Falkow, S. (1985) A single genetic locus encoded by Yersinia pseudotuberculosis permits invasion of

cultured animal cells by Escherichia coli K-12. Nature 317: 262-264. [2]Denise M. Monack,Julie A. Theriot(2001).” Actin-based motility is sufficient for bacterial membrane protrusion formation and

host cell uptake”. Cellular Microbiology (2001) 3(9), 633-647

Endosymbiosis between Eukaryotes and

Cyanobacteria(inv-llo +)

Agapakis CM, Niederholtmeyer H, Noche RR, Lieberman TD, Megason SG, Way JC, & Silver PA (2011).

Towards a synthetic chloroplast. PloS one, 6 (4) PMID: 21533097

CHO cells J774 macrophages

cells Invasin(inv) from Yersinia pestis

listeriolysin O(llo) from Listeria

monocytogenes

Cells:

• J774 macrophage cells line

• Induced pluripotent stem cells

SO2 H2S S CysI SQR

Our work

J774 macrophage cell line

with cyanobacteria invasion

Induced pluripotent stem cells(iPS cells)

with cyanobacteria invasion

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H2S SO2

Practical Applications To confirm our project

Practical Application →purifying polluted air →confirming our project practically

SO2 CO2

Practical Experiments to confirm: The Experiment Chung Hwa Pulp Corporation

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Co-culture:

Engineered bacteria+

industrial waste

For waste gas cleaning

Water layer Gas layer

Practical Applications: The Experiment

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WORK

NYMU-Taipei

H2S producing assay

For waste water cleaning

Water layer Gas layer

Practical Applications: The Experiment

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WORK

NYMU-Taipei

H2S producing assay

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H2S SO2

Completing whole parts!!

Solved!!

Synechocystis

PCC7942

Solved!!

Sulfite reductase

Solved!! sqr

Solved!!

INV-LLO

Synechocystis

PCC7942

Work!!

NYMU-Taipei

Integrating all factors!!

Solving all problems!!

Let’s go to VENUS!!

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Human Practice—what have they done Brand New Idea Communicating with aliens by Arecibo code Meeting with other Taiwan iGEM teams

6/1 Meeting with NTU-Taida

8/15 Taiwan iGEMer Fest (Press Release)

Having Fun

Going to Hot Spring Valley

Visiting Taipei Astronomical Museum

Advocating our project Board Game

iGEM song

iGEM short film

Advocating iGEM & synthetic biology

7/27 Synthetic Biology Worldwide

8/26 Welcome Party for Freshmen

9/13 Exhibition of iGEM in NYMU

Arecibo code scattering around Taipei

6/1 Meeting with NTU-Taida

8/15 Taiwan iGEMer Fest (Press Release)

Their E.coli

works

Creating a whole new method to exchange and

discuss ideas within Taiwan iGEM teams for themselves

and the future 2013、2014、2015… Taiwan iGEMers

Let people join iGEM in the future easier

Advocating 2012 NYMU

iGEM team

B o a r d G a m e i G E M s o n g

i G E M s h o r t f i l m

Advocating iGEM &

synthetic biology

7/27 Synthetic Biology Worldwide 8/26 Welcome Party for Freshmen

9/13 Exhibition of iGEM in NYMU

To attract more people to

What do those mean ?

To let more people to what is iGEM & synthetic biology

know

To make people iGEM &synthetic biology interested in

To let some of those people who start to realize iGEM &synthetic biology to us!! join

engage in synthetic biology field!

Achievements We have submitted 11 parts to part registry.

We have characterized and proved that these new biobrick parts:

SQR, CysI, DsrI really work as our expectation.

We have modified BBa_K299812 (INV+LLO) and elevated our

idea to a higher level, to create venusian!

We have held a meet-up with Taiwan iGEMers, with board

games, iGEM songs, movies and coorperated with NTU-

Taida.

We have already outlined and detailed an extremely new human

practice - outer space human practice project (New human

practice approaches!!).

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NYMU-Taipei

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We are the first iGEM team who tries to…

communicate with the aliens!!

We had lots of… FUN & FUN &FUN!!

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Reference

P1 http://www.google.com.tw/imgres?hl=zhTW&sa=X&tbm=isch&prmd=imvns&tbnid=qOU_cLnW5mjJKM:&imgrefurl=http://www.geekosystem.com/ben-stiller avataroscars/&docid=cztpwj_qRRdhBM&imgurl=http://www.geekosystem.com/wpcontent/uploads/2010/03/benstilleravatar.jpg&w=461&h=285&ei=97tnUOmbGM7umAX8hoCIAw&zoom=1&iact=rc&dur=375&sig=109169273201630242688&page=2&tbnh=139&tbnw=197&start=18&ndsp=22&ved=1t:429,r:12,s:18,i:167&tx=144&ty=61&biw=1280&bih=642

P5

computer fixerhttp://www.google.com.tw/imgres?hl=zh-TW&tbm=isch&tbnid=h2Q2KTB_ECeQFM:&imgrefurl=http://www.techsonsite.co.uk/&docid=REnxlPXOqynqOM&imgurl=http://www.techsonsite.co.uk/wp-content/uploads/2011/03/milton-keynes-computerrepairs.png&w=359&h=268&ei=Vb1nUNaWO4XLmgWCk4HICg&zoom=1&iact=hc&vpx=977&vpy=205&dur=18&hovh=194&hovw=260&tx=197&ty=124&sig=109169273201630242688&page=1&tbnh=142&tbnw=192&start=0&ndsp=18&ved=1t:429,r:11,s:0,i:101&biw=1280&bih=642

lightbulb

http://www.google.com.tw/imgres?hl=zhTW&tbm=isch&tbnid=kfPkZWWGCXwYbM:&imgrefurl=http://blog.enn.com/%3Fp%3D481&docid=MdxonwH6otD7aM&imgurl=http://blog.enn.com/wp-content/uploads/2010/10/lightbulb.jpg&w=315&h=387&ei=gL1nULjHFMXLmAX-8YGQDQ&zoom=1&iact=rc&dur=143&sig=109169273201630242688&page=2&tbnh=142&tbnw=113&start=23&ndsp=31&ved=1t:429,r:6,s:23,i:162&tx=64&ty=89&biw=1280&bih=642

47 NYMU-Taipei

http://www.google.com.tw/imgres?hl=zh-

TW&tbm=isch&tbnid=cgZK8mOqlDXOaM:&imgrefurl=http://xiaosinyi.pixnet.net/blog/post/33570223motivation!&docid=UrV5bFu-KDNZVM&imgurl=http://4.bp.blogspot.com/_ccEezkECpdg/TSSQQ_jaMHI/AAAAAAAADsA/H1sMe6pdUKA/s1600/motivation.jpg&w=900&h=900&ei=mb1nUPScF4_JmQXQ94CwCg&zoom=1&iact=rc&dur=1&sig=109169273201630242688&page=2&tbnh=139&tbnw=139&start=17&ndsp=24&ved=1t:429,r:19,s:17,i:186&tx=69&ty=83&biw=1280&bih=642

p7

relay race

http://www.google.com.tw/imgres?hl=zh-TW&tbm=isch&tbnid=98OqvAVMjU7UfM:&imgrefurl=http://thepauls.wordpress.com/2011/01/31/the-gospel-baton/&docid=2trO2aDES7n-fM&imgurl=http://thepauls.files.wordpress.com/2011/01/pass_the_baton.jpg&w=1024&h=768&ei=y71nUJTJAe_LmAWiiIDQCg&zoom=1&iact=rc&dur=137&sig=109169273201630242688&page=1&tbnh=134&tbnw=230&start=0&ndsp=16&ved=1t:429,r:3,s:0,i:76&tx=110&ty=74&biw=1280&bih=642

p8

book

http://www.google.com.tw/imgres?hl=zh-TW&tbm=isch&tbnid=VUrSo60jjoGlSM:&imgrefurl=http://www.armenianweekly.com/2012/04/06/the-armenians-and-the-book-exhibit-to-open-at-harvard/&docid=DNtmCdMCXvPmIM&imgurl=http://www.armenianweekly.com/wp-content/uploads/2012/04/book.jpg&w=600&h=450&ei=7L1nUO-oC6POmgWu-4GoBg&zoom=1&iact=rc&dur=279&sig=109169273201630242688&page=2&tbnh=139&tbnw=188&start=18&ndsp=24&ved=1t:429,r:5,s:18,i:144&tx=83&ty=53&biw=1280&bih=642

p9

shortcut

http://www.google.com.tw/imgres?hl=zh-TW&tbm=isch&tbnid=p3FTTaefkBrCuM:&imgrefurl=http://lostinexcel.blogspot.com/2012/02/all-worksheet-shortcuts-one-place.html&docid=_Zhh_o5KDcQe6M&imgurl=http://4.bp.blogspot.com/-0nKLyRIQYME/T0JGf7ScQpI/AAAAAAAAEHA/rnRE-MtXIFk/s1600/worksheet%252Bshortcut%252Bkeys.jpg&w=517&h=342&ei=EL5nUPzIB6WNmQXno4CgBg&zoom=1&iact=hc&vpx=748&vpy=173&dur=96&hovh=182&hovw=276&tx=121&ty=108&sig=109169273201630242688&page=4&tbnh=128&tbnw=193&start=69&ndsp=24&ved=1t:429,r:9,s:69,i:320&biw=1280&bih=642

p10

Arecibo code

48 NYMU-Taipei

http://www.google.com.tw/imgres?hl=zh-TW&tbm=isch&tbnid=udPyJF8_tSyg5M:&imgrefurl=http://pzzled.yolasite.com/and-everything-in-between.php&docid=KpEVCsQ7jEh26M&imgurl=http://pzzled.yolasite.com/resources/eg-arcibo-glyph.jpg%253Ftid_1258886625188&w=462&h=460&ei=S75nUML-GYGKmQXuwIGQAQ&zoom=1&iact=rc&dur=224&sig=109169273201630242688&page=1&tbnh=131&tbnw=132&start=0&ndsp=23&ved=1t:429,r:2,s:0,i:73&tx=94&ty=70&biw=1280&bih=642

p18

surprise

http://www.google.com.tw/imgres?hl=zh-TW&sa=X&tbm=isch&prmd=imvns&tbnid=HvNqWchw5GJZuM:&imgrefurl=http://dealcetera.com/sears-canada-72-hour-surprise-sale-october-8-10th-savings-and-discounts-up-to-50-off-searsca/&docid=7C58RkoEAUraJM&imgurl=http://dealcetera.com/wp-content/uploads/2009/10/surprise.jpg&w=267&h=400&ei=yb5nUOX-PM7UmAWq4IG4BQ&zoom=1&iact=rc&dur=250&sig=109169273201630242688&page=2&tbnh=141&tbnw=94&start=18&ndsp=26&ved=1t:429,r:7,s:18,i:151&tx=55&ty=54&biw=1280&bih=642

cat ball

http://www.google.com.tw/imgres?hl=zh-TW&tbm=isch&tbnid=xE69VYj-jkZ-oM:&imgrefurl=http://fineartamerica.com/featured/6891-pet-impressions-cat-with-ball-kyle-adamache.html&docid=KSw-QmydUtAoIM&imgurl=http://images.fineartamerica.com/images-medium/6891-pet-impressions-cat-with-ball-kyle-adamache.jpg&w=525&h=700&ei=6L5nUP-QNozMmAWJu4AY&zoom=1&iact=rc&dur=241&sig=109169273201630242688&page=2&tbnh=144&tbnw=117&start=18&ndsp=24&ved=1t:429,r:5,s:18,i:144&tx=73&ty=98&biw=1280&bih=642

join us

http://www.google.com.tw/imgres?start=163&hl=zh-TW&tbm=isch&tbnid=vEmiKOcs244jwM:&imgrefurl=http://km2.iiietc.ncu.edu.tw/xms/content/show.php%3Fid%3D29907&docid=y9V1AVfLMAvYFM&imgurl=http://km2.iiietc.ncu.edu.tw/xms/data/256/content/86E6DD72A5A058C0DB5D/attach/C1000086JoinUs.jpg&w=600&h=848&ei=Er9nUJuuCerKmgWSvoHYCQ&zoom=1&iact=hc&vpx=472&vpy=278&dur=1437&hovh=267&hovw=189&tx=119&ty=183&sig=109169273201630242688&page=8&tbnh=133&tbnw=94&ndsp=25&ved=1t:429,r:2,s:163,i:243&biw=1280&bih=642

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NYMU-Taipei