الفهرس 
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Abstract
from the present study, we can conclude that;
1. Deep water sample and squid are good sources for isolation of fluorescent bacteria.
2. Statistical optimization designed new culture medium for the highest quantity of GFP by Egyptian Pseudomonas gessardii E2 strain using RSM.
3. The optimized medium to obtain the highest GFP production has the following compositions (g/l); peptone, 6; yeast extract, 3; glycerol, 4; NaH2PO4, 1.14; sea water, 68.121%; 31.87 % distilled water.
4. Formation of GFP silver and gold-nanocomposite enhance the fluorescence intensity of green fluorescent protein.
5. This study confirmed the enhancement anticancer activity and drug delivery by formation of GFP with Ag-nanocomposite and Au-nanocomposite for treatment of Ehrlich Ascites Carcinoma (EAC).
6. Green fluorescence protein can be used in the imaging of malignant cells in mouse model.
7. GFP-Ag nanocomposite and GFP-Au-nanocomposite can be used in theranostic approach for assisting the transition from traditional, nonspecific medicine to modern precision treatment.
7. Recommendations
Our study recommend that:
1. Large scale production of GFP nanocomposites for medical applications.
2. Investigation of GFP silver and gold nanocomposites on other types of cancers.
3. Application of GFP-Ag-nanocomposite and GFP-Au-nanocomposite in cancer theranostic application and other applications like biosensors.
8. Summary
The obtained results can be summarized as the following:
1. Eleven fluorescent bacterial isolates were isolated from squid and water samples (seven fluorescent bacteria were isolated from different parts of squid and four fluorescent bacteria from sea water) using five different isolation media.
2. Out of five isolation used media, sea water complete agar medium provided the highest growth of fluorescent bacteria.
3. Four bacterial isolates were selected based on the primary screening by visual scoring for fluorescence production.
4. from the secondary screening results, E2 bacterial isolate was selected as the highest fluorescence bacteria based on fluorescence intensity and green fluorescent protein concentration and purity.
5. E2 isolate as the most potent fluorescent bacteria was identified based on its morphology as gram -ve rod bacilli with single polar flagella.
6. Based on its biochemical characterization, E2 isolate was identified as Pseudomonas fluorescence with a probability percentage of 95%.
7. Genetically, E2 isolate was identified as Pseudomonas gessardii and it was submitted to GenBank under accession number OQ285875.
8. The environmental conditions for the highest growth and fluorescence intensity of Pseudomonas fluorescence E2 strain was inoculum period for 30hrs under static state at 25°C and pH 7.5.
9. The significant factors affecting the fluorescence production of Pseudomonas gessardii E2 strain was peptone, sea water, glycerol, and sodium dihydrogen phosphate.
10. The optimum concentration of the significant factors was peptone, 6 g/l; glycerol,4 g/l; NaH2PO4,1.14 g/l and sea water, 68.121% that increase the fluorescence intensity more than two-fold compared to the basal medium.
11. The GFP was purified with 96 % purity.
12. Silver nanoparticles was biosynthesized using Fusarium oxysporum as a brownish solution with absorption peak at 410 nm and zeta potential of -34.6 (mV).
13. Gold nanoparticles was biosynthesized as a purple-colored solution with absorption peak at 530 nm and zeta potential of -31.6 (mV)
14. The ideal formed GFP nanocomposites was at 3:1 ratio of GFP to nanoparticles which enhanced the fluorescent intensity of GFP.
15. TEM images of the ideal formed GFP-nanocomposites showed the GFP coating silver nanoparticles of size range from 23 to 25nm and gold nanoparticles from 17 to 19nm.
16. Based on in vivo anticancer efficacy study of nanoparticles and GFP composites on Ehrlich solid carcinoma, it was found that GFP composites decreased the tumor size compared with nanoparticles and especially for gold nanocomposite.
17. GFP-Au-nanocomposite recorded the lowest level of Malondialdehyde (MDA) (1.2 nmol/mg/ protein) and the highest level in Superoxide dismutase enzyme (SOD) (3.3 nmol/mg/ protein) in Ehrlich solid carcinoma bearing mice.
18. For Immunological studies, the lowest levels of Interleukin-1 beta and Tumor Necrosis Factor- α was recorded by GFP-Au-nanocomposite which gave 63.3 and 98.3 pg/mg/protein, respectively.
19. Histopathological studies in mice models illustrated that the GFP-Au-nanocomposite showed the highest decrease in neoplastic cells that appeared as small foci surrounded by necrotic tissue.
20. Diagnostic value of GFP imaging for Ehrlich tumor in mice revealed that Live tumor cells showed a strong fluorescence intensity whereas necrotic cells almost completely lose their fluorescence intensity.