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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
Over expression of human interferon α 2a gene in Chlamydomonas reinhardtii under a biotic stress
2
Emad Hamdy, M.Y,
2
I M El-Manawy,
3
Rashad Kebeish
1, 3
Sherif Nasshy,
and
1
EL-Ayouty
1
Botany Department, Faculty of Science, Zagazig University, Egypt
2
Botany Department, Faculty of Science, Suez Canal University, Egypt
3
Clinical Pathology Department, Kaser El-Aini Medical School, Cairo University, Egypt
The Interferon- α2a (IFN- α2a) is a principle cytokine that has regulatory role in mammalian immune systems. The coding
sequence of human interferon α2a(hu-IFN-α2a) gene was cloned into binary vector pTRAK, a derivative of pPAM(gi13508478).
The recombinant plasmid was transferred into Agrobacterium tumefaciens strain (GV3101). Then the hu-IFN-α2a gene was
introduced into Agrobacterium-mediated transformation to Chlamydomonas reinhardtii (Kumar
et al.
2004)and (Cha
et al.
2012), with modifications. carrying hu-IFN-α2a gene. to confirmed the well integrity and positive colonies. ten cell line were
taken for PCR amplification. Results indicate that hu-IFN-α2a gene has been integrated into the Chlamydomonas reinhardtii
genome, with transcription products of. hIFN-α2a. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE)
manifested a specific protein band at 19.5 KD of five different transgenic Chlamydomonas reinhardtii. Hu-IFNα2A protein
was detected by the enzyme linked immunosorbant assay (ELISA) and RT-PCR. These results suggest that Chlamydomonas
reinhardtii can be used to produce many biologically active mammalian proteins that accumulate in target organelles such
as protein bodies with low cost with high efficiency. Cytotoxicity of hu-IFNα2a extracts from three line of transgnic alga was
evaluated through intrapertioneal and subcutaneous studies using Albino mice infected with Ehrlich ascites carcinoma. The
results revealed that an obvious reduction in fluid volumes by (85%,68% and 75%less than untreated animals as the infected
mice injected intraperitoneal with 21.45 ug\kg body wt. Whereas under subctaneouus injection the origenal mass size reduced
by 93% as compared with untreated animals.
Keywords:
Human interferon α2a,
Agrobacterium tumefaciens
, SDS-PAGE, TAP medium, immunosorbant assay (ELISA),
Ehrlich ascites carcinoma (EAS CELLS)
Biography
Emad hamdy is currently a permanent researcher at Biotechnology Labs, Faculty of Science, Ismailia, A Suez Canal Univeristy,I has Bachelor science in Chem-
ist-Botany(good grade)- Faculty of Science- Zagazig- university- Sharkia – Egypt, Diploma in Applied Microbiology (good grade) - Faculty of Science - Banha-
Univerisity. and MSc in Microbiology (2014), Faculty of Science, Zagazig University, Egypt. I accumulated 12-years of experience in quality and infection control,
and appointed to a number of key jobs; ISO 9001:2015 Lead Auditor, quality manager and quality and infection control manager in Techno scan group, and senior
quality assurance specialist and internal auditor at NSA Company, 2010- till date.
em.emad86@yahoo.comEmad Hamdy et al., J Ecosys Ecograph 2018, Volume: 8
DOI: 10.4172/2157-7625-C2-035