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bold'> which regulates the color of the culture and expression of CsgA-ECs.</span><o:p></o:p></span></p> | bold'> which regulates the color of the culture and expression of CsgA-ECs.</span><o:p></o:p></span></p> | ||
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Revision as of 19:01, 18 September 2015
Background and overview
Overview
Background
In 1930s, a new, strange disease happened in
Toyama village, Japan, called Itai-Itai disease, whose symptoms were exactly
the same as it was. Patients had insufferable pains in bones. The whole body
anywhere was very easy to fracture. They had been shouting “Pain!
Pain!” until death. Now we know, the cause of the disease is cadmium poisoning.
The river that local people used for drinking and irrigating crops was polluted
by the sewage, containing a large amount of cadmium. It was discharged from the
Zinc smelting plant, which was built on the upper reaches of the river. Cadmium
through food chain gradually enriched in human body. Up to now, in 2014, a
village in Guangxi, China also has the occurrence of Itai-Itai disease.
Figure1: Patients with bone deformation due
to cadmium poisoning
Figure
2: A child was moaning because the great pain caused by Itai-Itai disease.
Cadmium’s entering human body can make it a
huge loss of calcium in bones and trigger a series of damage, like
osteoporosis, bone atrophy, joint pain and so on. Once had a patient, hit a
sneeze, unexpectedly leading to several fracture. Even mild cadmium poisoning
can cause lung and kidney damage, like toxic pulmonary edema and chemical pneumonia.
Figure 3:
Cadmium does harm to the bone seriously.
Asia is the world's largest primary cadmium
metal producing areas, mainly in China, South Korea, and Japan. But industry that is related to cadmium
is globally existed:Smelting, spraying, welding and casting bearing surface of cadmium;Stope;Rods in nuclear reactors;Neutron absorber;Electroplating factory, and
so on.
So, do you think cadmium is far
away from our daily life? No.
In June 2010, the U.S. Consumer Product
Safety Commission confirmed that the glasses sold by McDonald were found to
contain cadmium elements in the paintcoat, which could exude from the surface of the glasses. About
twelve million glasses were recalled in the United States.
Figure
4 :The paintcoat
of the glasses sold by McDonald were detected to contain cadmium elements
In 2012,the report from the environmental
organization of Canada about Potential Damage of Heavy Metals in Cosmetics
said, almost all of the tested cosmetics contain toxic substances, including
cadmium.
In June 2012,Shandong Province, Rizhao
City inspection and Quarantine Bureau
seized 24 tons of heavy sauries from Japan , whose Cd content exceeded
the safety standard.
Figure
5:Rizhao(China) inspection and quarantine bureau staff at the time of
sampling inspection, found that cadmium content of these 24tons saury exceeded
the safety standard.
Smoking 20 cigarettes a day, 2 to 4
micrograms of cadmium can be taken in
Figure 6:Cadmium containing rice due to water and soil pollution.
Since there is no effective treatment of Itai
Itai disease, and cadmium stored in the body did not rule out in a safe and
effective method. Therefore, it is especially important to eliminate cadmium
pollution on the environment; this is the fundamental measure to prevent the
occurrence of pain!
So we got the idea of our project: Super cadmium ion killer
Project
Overview
Concerning
cadmium pollution, the iGEM team 2015 SCUT would
like to present super cadmium ion killer, a bacterial platform designed to
detect and absorb the cadmium in sewage all over the world. We employ E.coli
superficial fiber curli protein—CsgA and synthetic phytochelatins (ECs) to form
CsgA-ECs, which can effectively remove cadmium ion in wastewater. Detection of cadmium ion presence in the sewage and
indication the concentration with various colors form the first part of the
super killer system. Expression of CsgA-ECs coupled to the sensor for
cadmium ion forms the second part of the super killer system. When our E.coli is
in a certain condition of low cadmium ion concentration, only the green chromoprotein cjBlue will be
expressed, leading to a color of green in culture. On the contrary, the
organism is on red alert with high concentration cadmium ion, producing a red fluorescent protein RFP as well as CsgA-EC(s) to absorb the ion.
It would be carried out until the concentration of cadmium ion dropping to a
low level. And the organism comes back to the origin state, the color return to
green again. As described above, our super cadmium ion killer system is
efficient, economical and real-time monitoring, which has great potential
application value in the treatment of cadmium pollution in the environment.
Figure
7: Super cadmium ion killer absorbing process. With the presence of cadmium
ion, the organism expresses CsgA-ECs to absorb the ion and turns red from
green.
Brief
introduction
l cadmium
killing
Cuili
fibers are powerful amyloid protein nano fibers, with a radius about 4-7 nm,
which are located in the surface of bacteria and expand as floccules. Assembled
by 13KDa CsgA extracellularly, Curli are ideal gene engineering application
platforms for extracellular absorption. Synthetic phytochelatins (EC) are naturally
occurring thiol-rich peptides with primary structures (Glu-Cys) n-Gly (n: repeat
unit), effectively binding Cd2+ and Hg+. Based on the
features described above, our project aim at assemble a device for removing
cadmium by fusing CsgA and ECs to achieve absorbing Cd2+ extracellularly.
•
Cd2+
monitoring (sensor system)
The
mercury-sensing regulatory protein, MerR, which regulates mercury resistance
operons in Gram-negative bacteria, was subjected to directed evolution in an
effort to generate a MerR mutant that responds to Cd but not Hg. MerR binds cadmium ion, when it presences in the culture. This provokes an allosteric
change in the MerR protein that triggers unwinding of the spacer DNA, allowing
transcription initiation of the mer operon, which regulates the color of the culture and expression of CsgA-ECs.