Projects.html
<!DOCTYPE html>
Welcome To Wiki Page
WATER IS SOUL OF UNIVERSE, A SOUL THAT FILLS LIFE, LOVE AND HOPE IN THIS WORLD, A SOUL WHICH HAS EVENTUALLY FADED INTO DARK CORNERS, A SOUL WHICH IS NOW CRYING FOR ENLIGHTMENT, A SOUL WHICH NEEDS AWAKEMENT WITHIN ALL OF US!!!
It is not just a molecule of oxygen and hydrogen, it is plethora of emotions and every day struggle. So water the essential requisite of nature, within which life originates and evolve, has now been completely intoxicated and destroyed by varied human activities and practices. We human in this metamorphism of evolution, have devastatingly affected this natural lineage and now the biggest problem posed by these un-regulated, and un-monitored human activities is release of heavy metal as waste product in water–bodies, as industrial discharge. Which increases concentration of these metals beyond permissible limits.
Thus, heavy metals (trace metals) becomes one of the most persistent pollutants in wastewater. The discharge of high amounts of heavy metals into water bodies leads to several environmental and health impacts, like –
- Death of aquatic life
- Destruction from sedimentation
- Algal blooms
- Habitat destruction
SOURCES OF VARIOUS HEAVY METALS
| HEAVY METAL | SOURCES |
|---|---|
| Arsenic(As) |
|
| Mercury(Hg) |
|
| Chromium(Cr) |
|
| Lead(Pb) |
|
| Cadmium(Cd) |
|
| Nickel(Ni) |
|
| Manganese(Mn) |
|
| Copper(Cu) |
|
| Zinc(Zn) |
|
Biological remediation processes (microbial remediation and phytoremediation) are indicated to be very effective in the treatment of heavy metal pollutants in wastewater. The expression of metal-binding proteins or peptides in microorganisms and plants in order to enhance heavy metal accumulation and/or tolerance has great potential. Several different peptides and proteins have been explored. The overexpression of metal binding proteins has been widely exploited to increase the metal binding capacity, tolerance or accumulation of bacteria and plants.
So, we as “TEAM DEI” are working in direction to reclaim this polluted water, by preparing a bio-chip(overexpression of metal binding protein), which removes heavy metal toxicity from industrial effluents, thereby making this water re-usable for activities like agriculture practices, sanitation purposes, and drinking; hence the project is titled-:
“Bio-Beads for removing heavy metal toxicity from industrial effluents“
The overexpression of metal binding proteins has been widely exploited to increase the metal binding capacity, tolerance or accumulation of bacteria and plants. An expression level of transgene/gene of interest depends upon strength of promoter. Architecture of these promoter elements can be redesign for modifying the gene expression. The basic rationale behind developing such modified promoters lies in the notion that the transfer of the upstream DNA sequence/cis-element that binds a specific trans-factor from one promoter into a different promoter containing the TATA sequence might result in a novel regulatory or transcription model. Hereby in our project we are focusing on overexpressing the modified Top-4 metal binder protein (modified iGEM part BBa¬_K1478002) and Human Metaloprotein- 3 (HMP3) by custom synthesizing strong promoter suitable for expression in microbes and plants. In our previous studies strong promoters had been developed. We would also fabricate a bio-chip by immobilizing those bacteria strongly expressing modified Top-4 metal binder protein (iGEM part BBa¬_K1478002) and HMP3 for fast, easy and user handy purification of industrial waste water which can be further usable for irrigation purpose.
“AIM“
All over the world large populations are utilizing potable water contaminated with varied heavy metals like arsenic (As), lead (Pb), cadmium (Cd), cobalt (Co) etc. with concentrations much higher than the permissible levels. Rapid industrialization in the field of mining, surface finishing, energy and fuel producing, fertilizers, pesticides, metallurgy, iron and steel, electroplating, electrolysis, electro-osmosis, leather, photography, electric appliances manufacturing, metal surface treating, aerospace and atomic energy installations, wastes containing metals are released in the environment and become available as potent contaminants. Industrial effluents possess high heavy metal toxicity which is a source of many hazardous diseases such as high exposure of cadmium can lead to obstructive lung diseases and has been linked to lung cancer. Cadmium may also cause bone defects in humans and animals. High doses of copper can cause anaemia, liver and kidney damage, and stomach and intestinal irritation. People with Wilson's disease are at greater risk for health effects from overexposure to copper. Exposure to high levels of arsenic can cause death. All types of arsenic exposure can cause kidney and liver damage. Hence to control the heavy metal toxicity of cadmium, copper, zinc, arsenic we are aiming to develop a microbial based biochip having overexpressed heavy metal binder protein for decreasing the toxicity of industrial effluents.
PROJECT IN DETAIL
(A) MATERIAL =>
Materials (list the pre-existing iGEM parts used, use of iGEM parts is mandatory)
| Synthetic Cis elements | Custom Synthesized |
|---|---|
| MMV Core promoter & Enhancer | Available at team leader |
| Part: BBa_K1478002 | IGEM part |
(B) METHOD =>
STEP 1
Designing of synthetic Promoter:
Promoters consist of several discrete domains arranged in a specific orientation and are modular, such that redesign of the ‘cis-architecture’ of a promoter DNA across its sequence backbone is feasible (Gurr and Rushton 2005; Rushton et al. 2002). The position/s of important functional domain/s between two different promoters can be shuffled/ exchanged (inter-/intra-molecularly), preferably upstream of the TATA box, to generate a unique transcriptional module performing the functions of both donor and acceptor (Acharya et al. 2014; Comai et al. 1990; Patro et al. 2012; Ranjan et al. 2012). The resultant functionality of the newly derived synthetic module depends upon the altered positioning of discrete cis motif/s with respect to their positions, spacing, orientation and copy number/s (Fessele et al. 2002; Tjian and Maniatis 1994).
In the earlier studied the GFP fluorescence of transformed E. coli with CaMV, MMV and FMV promoter constructs in presence of GFP reporter gene was estimated by confocal laser scanning microscopy. Ratios of activities between pPMMVFLt12GFP/ pPCaMV35SGFP and pPFMVSgt3GFP/pPCaMV35SGFP were found to be 12.9 and 5.06, respectively. The activities of these promoters in presence of GUS reporter gene were also determined biochemically and the ratios of activities were found to be 14.08 and 4.88, respectively.
As the maximum activity was observed in pPMMVFLt12 construct, for the present study the strong promoter having the synthetic cis elements upstream will be designed using bioinformatics tools
STEP 2
Construct Formation:
- Cloning of synthetic promoter in PUC119:- The custom synthesized cis element is cloned upstream of core promoter of puc119 in cloning Vector.
- Hybridizing synthetic promoter:- The developed construct will be hybridized with IGEM part.
STEP 3
Entrapment of the cells:
Microbial whole cells are efficient, ecological, and low-cost catalysts that have been successfully applied in the pharmaceutical, environmental, and alimentary industries, among others. Microorganism immobilization is a good way to carry out the bioprocess under preparative conditions. The main advantages of this methodology lie in their high operational stability, easy upstream separation and bioprocess scale-up feasibility.
The developed construct will be transformed in E.coli cells and positive recombinants E.coli cells will be entrapped in sodium alginate and hydrogels.
STEP 4
Efficacy testing of biochips:
Automatic Absorption Spectrophotometer will be opted for detection of heavy metals before and after treatment of the wastewater with developed biochip.
