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Journal of Ecosystem & Ecography | ISSN: 2157-7625 | Volume: 8

June 28-29, 2018 | Alexandria, Egypt

International Pre Conference Workshop on

Microbial Ecology & Eco Systems

Role of metallothionen and glutathione-s-transferase expression in

Chlamydomonas reinhardtii

for

detoxification sewage water from some heavy metals

Abdelhamed M Elkbash, EL-Ayouty Y.M., Rashad Kebeish, Samy

and

M M

Botany Department, Faculty of Science, Zagazig University, Egypt

T

he maintenance of ion detoxification in living cells is a fundamental physiological requirement for sustainable their

growth, development, and production. Metallothioneins (MTs) are a superfamily of cysteine-rich, low-molecular-weight

metalloproteins that bind heavy metal ions. These cytosolic metallopeptides are widely distributed in living organisms and

are thought to be involved in metal homeostasis, metal detoxification, and oxidative stress protection. Here in Transgenic

Chlamydomonas reihardtii

was constructed via cloning and sub cloning with PTRA-GST and PATRA-PT- Smta plasmds.

The first one was restricted via ASC1 and BAM and the second restricted by BAM. Their products were purified according to

protocol kits and to allowed to ligase. The product was introduced into Agrobacterium-mediated, and then transformed to

Chlamydomonas reinhardtii. The transgenic alga was selected on solid TAP medium supplemented with Basta at concentration

(12ug\ml). The efficiency of algal trasformation was confirmed by SDS –gel electrophoresis and RT- PCR. To evaluate that

toxicity of heavy metal an experiment was conducted in which the wild and transgenic

Chlamydomonas reinhardtii

allowed

to grow in sewage water for six days and the rate of removal heavy metal with calculated. Results revealed that, the transgenic

Chlamydomonas reinhardtii

has MtIII clearly can have an important role in metal detoxification. Transgenic

Chlamydomonas

reihardtii

with metallothionen gene was selective for sequestration of cytosolic Cd2

+

. It has been found that MtIII synthesis is

related to degree of pollution in an aquatic environment.

Keywords:

Autophotosynthetic, biomarker, Cd2

+

,

Chlamydomonas reihardtii,

glutathione-s-transferase, heavy metals,

metallothionein.

Biography

Abdelhamed M Elkbash is currently a researcher at Biotechnology Labs Faculty of Science, Zagazig University, Egypt. He has Bachelor of Science, Alazher

University, Microbiology Department. His scientific grade (Excellent). He works as a Biologist in the Central Lab. for drinking water, Behira Water and Drainage

Company (BWADC), where he is responsible for water and wastewater treatment analysis (chemical and microbiological) for surface and ground water. He

accumulated 13-years of experience in different Labs, and implementation of ISO 17025 (general requirement for competence of testing and calibration laboratories),

and ISO 22000 (food safety management system) applicable for safety of drinking water.

Elkabash2020@yahoo.com

Abdelhamed M Elkbash et al., J Ecosys Ecograph 2018, Volume: 8

DOI: 10.4172/2157-7625-C2-035