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03716nam a22003737a 4500 |
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1511212 |
| 041 |
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|a eng
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| 100 |
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|9 546568
|a المصطفى، ناريمان عبدالكريم سلمان
|e مؤلف
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| 245 |
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|a إنتاج ودراسة خصائص أجزاء من مستقبل ال EGFR البشري (L2 و CR2) على شكل بروتينات مركبة في بكتيريا BL21وEscherichia Coli Origami B
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| 246 |
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|a Expression and Characterization of Human EGFR L2 and CR2 Domains Using the Escherichia Coli Origami B and BL21 Strains
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| 260 |
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|a المنامة
|c 2017
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| 300 |
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|a 1 - 76
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| 336 |
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|a رسائل جامعية
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| 502 |
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|b رسالة ماجستير
|c جامعة الخليج العربي
|f كلية الدراسات العليا
|g البحرين
|o 1406
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| 520 |
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|a The Epidermal growth factor receptor (EGFR) is a cell surface protein composed of an extracellular ligand-binding domain, transmembrane segment, and an intracellular tyrosine kinase domain. The EGFR plays a key role in many of the cellular processes (cell proliferation, migration) involved in cancer development and has been proven to be one of the most important and rational target for anti-tumor strategies. Monoclonal antibodies (mAbs) bind to the extracellular domain of the EGFR and inhibit ligand binding to the receptor, inducing receptor dimerization and down regulation. Therefore, recombinant mAbs specifically targeting mutation in the extracellular domain of the EGFR would robustly enhance anti-tumor action and may perhaps become valuable therapeutic reagents. In this project, we used the Escherichia coli (E.coli) system of recombinant protein expression to produce the human native EGFR extracellular domains L2 and CR2. Synthetic L2 and CR2 sequences were cloned into pGEX4T-1 plasmid under the control of tac promoter. Two different Escherichia coli strains: Origami B and BL21 strains were used to produce the recombinant proteins each fused to Glutathione-S-Transferase (GST). Recombinant protein expression was analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and western blot assay. Our results show that the recombinant EGFR L2 and CR2 domains are expressed as soluble recombinant proteins. However, approximately 60% of the protein produced is trapped in inclusion bodies. The amount of soluble EGFR L2 and CR2 domains is higher in E.coli BL21 strains (35 %) cultured at 30ᵒC and induced using 1 mM IPTG. The soluble recombinant EGFR L2 and CR2 domains expressed in BL21 can be produced in a preparative scale, purified, and used to carry out structure function analysis and to develop functional monoclonal antibodies to EGFR.
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| 653 |
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|a البلازما
|a البروتينات المركبة
|a EGFR اللبشري
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| 700 |
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|9 546572
|a Fathallah, Mohammed Dahmani
|e Advisor
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| 700 |
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|9 546574
|a Hachemi, Sonia Bouguiba
|e Advisor
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| 856 |
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|u 9812-007-001-1406-T.pdf
|y صفحة العنوان
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| 856 |
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|u 9812-007-001-1406-A.pdf
|y المستخلص
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| 856 |
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|u 9812-007-001-1406-C.pdf
|y قائمة المحتويات
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| 856 |
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|u 9812-007-001-1406-F.pdf
|y 24 صفحة الأولى
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| 856 |
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|u 9812-007-001-1406-1.pdf
|y 1 الفصل
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| 856 |
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|u 9812-007-001-1406-2.pdf
|y 2 الفصل
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| 856 |
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|u 9812-007-001-1406-3.pdf
|y 3 الفصل
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| 856 |
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|u 9812-007-001-1406-4.pdf
|y 4 الفصل
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| 856 |
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|u 9812-007-001-1406-5.pdf
|y 5 الفصل
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| 856 |
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|u 9812-007-001-1406-6.pdf
|y 6 الفصل
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| 856 |
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|u 9812-007-001-1406-O.pdf
|y الخاتمة
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| 856 |
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|u 9812-007-001-1406-R.pdf
|y المصادر والمراجع
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| 856 |
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|u 9812-007-001-1406-S.pdf
|y الملاحق
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| 930 |
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|d y
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| 995 |
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|a Dissertations
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| 999 |
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|c 1011550
|d 1011550
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