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dc.contributor.advisor El Dorry, Hamza El Said, Nadine Hosny
dc.creator El Said, Nadine Hosny 2012-02-04T15:49:00Z 10000-01-01T15:49:00Z 2011 Fall 2012-02-04T15:49:00Z
dc.description.abstract The Atlantis II deep is the largest brine pool in the Red Sea of around 60 km2. It is located in the central Red Sea (21° 21' N., 38° 04' E.) at a depth of 2200 meters from the sea surface. The lower part of this pool, the lower convective layer (LCL), has extremely harsh environmental conditions characterized by a temperature of up to 68°C, pH 5.3, saturated salt (252 psu), anoxic condition, and high concentration of heavy metals. These harsh abiotic factors exert strong selective pressure in favor of microbial communities adapted to survive in such environment. Therefore, proteins and enzymes involved in the metabolic pathways and physiological processes of these microorganisms should have novel structural and enzymatic activities that could be of interest in biotechnological applications. In this work we utilized an integrated metagenomic approach to identify sequences of potential secreted subtilisin-like serine proteases in the LCL metagenomic datasets, and also functionally screened the LCL metagenomic fosmid library using skimmed milk (casein) as a substrate. Out of 1,167,000 LCL reads (average length 400 nucleotides/read), we obtained 286 sequences that matched potential secreted subtilisin-like serine proteases. The 286 matched reads were assembled into 31 contigs. In the functional analysis, we screened 10,656 clones that provided 9 positive candidates as judged by characteristic clear halo around the positive colonies. Pyrosequencing of 2 of the plates harboring functionally positive clones provided 5 open reading frames that matched subtilisin-like serine protease. This work provided a pivotal step toward identification of subtilisin-like serine proteases with novel catalytic activities. en
dc.description.sponsorship I would like to thank Professor Dr Hamza El Dorry (My advisor), Dr Mohammed Ghazy who co-advised and helped me out throughout the lab work. I would also like to thank Dr Ari Ferreira, Mr Hazem Sharaf and Mr Amged Ouf, for being a friendly helping hand whenever needed. I would also like to thank Professor Dr Rania Siam for her guidance and aid during the whole period I spent at AUC for completing my MSc. I extend my gratitude to everybody who provided me with the opportunity to work at KAUST during Summer 2010: Professor Dr Vladimir Bajic (for helping and advising me during the internship at KAUST) Dr Adam Dawe, Dr Intikhab Allam, Mr Allan Kamau and everybody at the computational Biology Research Center (CBRC). I would also like to thank KAUST for providing us with the funds to finish this Project. I would like to send my extreme love and appreciation to thank my family who has helped me out throughout my entire life to reach my goals: my father, mother, sister, and husband. Would also like to dedicate this work to my beloved Son baby Hamza who has witnessed lots of the hard work since he was still in my womb. And finally I would like to thank all people who have helped me finish this work in its best form, all my professors at AUC and Ain Shams University, colleagues and family. en
dc.format.medium theses en
dc.language.iso en en
dc.rights Author retains all rights with regard to copyright. en
dc.subject Red Sea en
dc.subject Subtilisin en
dc.subject Serine proteases en
dc.subject Metagenomics en
dc.subject Function based screening en
dc.subject Sequence based screening en
dc.subject Atlantis II Brine pool en
dc.subject.lcsh Thesis (M.S.)--American University in Cairo en
dc.subject.lcsh Biotechnology -- Red Sea -- Egypt.
dc.subject.lcsh Microbiology -- Red Sea -- Egypt.
dc.subject.lcsh Genetic engineering -- Red Sea -- Egypt.
dc.subject.lcsh Molecular biology -- Red Sea -- Egypt.
dc.title Identification of potential subtilisin-like serine proteases from the microbial community of the Red Sea Atlantis-II brine pool using a metagenomic approach en
dc.type Text en
dc.subject.discipline Biotechnology en
dc.rights.access This item is restricted forever en
dc.contributor.department American University in Cairo. Dept. of Biology en
dc.embargo.lift 10000-01-01T15:49:00Z

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  • Theses and Dissertations [1633]
    This collection includes theses and dissertations authored by American University in Cairo graduate students.

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