Additional file 3: Figure S3. Digestion of pHT01 plasmid and the amplified kIspS fragment with Ba... more Additional file 3: Figure S3. Digestion of pHT01 plasmid and the amplified kIspS fragment with BamHI and XbaI to 11 generate pHT01-kIspS construct. Marker: 2 log DNA ladder (1.0â 10.0 kb) NEB catalogue #N04695, 12 Lane 1: pHT01 (7.9 kb) digested by BamHI and XbaI, Lane 2: amplified kIspS fragment (1.7 kb) 13 digested by BamHI and XbaI.
Additional file 2: Figure S2. Colony PCR results of the recombinant plasmid pET28b-kIspS-C term i... more Additional file 2: Figure S2. Colony PCR results of the recombinant plasmid pET28b-kIspS-C term in BL21 (DE3) cells. 7 Marker: 2 log DNA ladder (1.0â 10.0 kb) NEB catalogue #N3200, Lane 1, 2 & 3 are positive results for 8 the colony PCR of pET28b-kIspS-C terminal in BL21 (DE3) cells.
Apigenin is one of the most studied flavonoids and is widely distributed in the plant kingdom. Ap... more Apigenin is one of the most studied flavonoids and is widely distributed in the plant kingdom. Apigenin exerts important antioxidant, antibacterial, antifungal, antitumor activities, and anti-inflammatory effects in neurological or cardiovascular disease. Chalcone isomerase A (chiA) is an important enzyme of the flavonoid biosynthesis pathway. In order to enhance the apigenin production, the petunia chi A gene was transformed for Astragalus trigonus. Bialaphos survived plants were screened by PCR, dot blot hybridization and RT-PCR analysis. Also, jasmonic acid, salicylic acid, chitosan and yeast extract were tested to evaluate their capacity to work as elicitors for apigenin. Results showed that yeast extract was the best elicitor for induction of apigenin with an increase of 3.458 and 3.9 fold of the control for calli and cell suspension culture, respectively. Transformed cell suspension showed high apigenin content with a 20.17 fold increase compared to the control and 6.88 fold m...
3 Abstract: Influenza A viruses infects birds, animals and humans and it has the ability to mutat... more 3 Abstract: Influenza A viruses infects birds, animals and humans and it has the ability to mutate and develop mutant variants which may cause unexpected events. There is an urgent need for new medications to combat influenza pandemics. Using the genome sequences of the influenza A virus performed previously, we designed and performed a combinatorial exhaustive systematic methodology for optimal design of universal therapeutic short-interfering RNA (siRNA) molecules targeting all diverse of the virus strains. By using the bioinformatics tools, we designed siRNA targeting influenza A virus polymerase genes. The selection and validation of optimal siRNAs was also studied. The influenza A virus subtype H1N1 and H5N1 were propagated in MDCK cells. TCID and HA assays were performed and real time PCR for assessment of viral load was used. Time- 50 dependent and dosage-dependent usage were performed and showed high decrease in viral load.
Plant defensins are small cysteine-rich peptides which belong to a group of pathogenasis related ... more Plant defensins are small cysteine-rich peptides which belong to a group of pathogenasis related defense mechanism proteins. The proteins inhibit the growth of a broad range of microbes and are highly stable under extreme environmental stresses. Tomato cultivation is affected by fungal disease such as Fusarium wilt. In order to overcome fungal damages, transgenic tomato plants expressing the Medicago sativa defensin gene MsDef1 under the control of the CaMV 35S promoter were developed. The Fusarium-susceptible tomato (Lycobersicum esculentum Mill) cultivar CastleRock was used for transformation to acquire fungal resistance. Hypocotyl with a part of cotyledon (hypocotyledonary) for young tomato seedlings were used as an explant material and transformation was performed using the biolistic delivery system. Bombarded shoots were selected on regeneration medium supplemented with hygromycin and suitable concentrations of BA, zeatin ripozide and AgNO(3). Putative transgenic plantlets of T(0) were confirmed by PCR analysis using primers specific for the transgene and the transformation frequency obtained was 52.3%. Transformation and transcription of transgenes were confirmed in T(1) by PCR, Southern hybridizations, and reverse-transcription PCR (RT-PCR). The copy numbers of integrated transgene into tomato genome ranged between 1-3 copies. Greenhouse bioassay was performed on the transgenic T(1) and T(2) young seedlings and non-transgenic controls by challenging with a vigorous isolate of the fungal pathogen Fusarium oxysporum f. sp. Lycopersici. The level of fungal infectivity was determined using RT-PCR with tomatinase specific primers. Transgenic lines were more resistant to infection by fusarium than the control plants. These results indicated that overexpressing defensins in transgenic plants confer resistance to fungal pathogens.
Abstract: Tuesday the 25 th of January is an enshrined date in Egypt, it will be always remembere... more Abstract: Tuesday the 25 th of January is an enshrined date in Egypt, it will be always remembered as the date Egyptians decided to fight for their freedom. Young Egyptians took to the streets and main squares. Their gathering ignited the revolution as three days later millions of protesters further gathered to demand changing the regime. They asked for “freedom and human dignity”. However, their demands rose to the resigning of Mubarak after unnecessary use of force by the police. Eighteen days later Mubarak stepped down ...
In Africa, six million children die from malnutrition before their fifth birthday each year (Worl... more In Africa, six million children die from malnutrition before their fifth birthday each year (World Bank statistics). More than 800 million people go to bed hungry, 300 million of them are children. “Simply because my people are hungry, that is no justification to give them poison.” Small words that were spoken by President Levy Mwanawasa of Zambia, who refused to accept milled grains from the United States during the worst food shortage crisis that Africa ever faced in 2002. Amazingly, African leaders of Zambia, Zimbabwe, Malawi and ...
In Africa, six million children die from malnutrition before their fifth birthday each year (Worl... more In Africa, six million children die from malnutrition before their fifth birthday each year (World Bank statistics). More than 800 million people go to bed hungry, 300 million of them are children. “Simply because my people are hungry, that is no justification to give them poison.” Small words that were spoken by President Levy Mwanawasa of Zambia, who refused to accept milled grains from the United States during the worst food shortage crisis that Africa ever faced in 2002. Amazingly, African leaders of Zambia, Zimbabwe, Malawi and ...
Abstract: Amflora provides a plausible reason for commercially cultivation of GM Crops in Europe.... more Abstract: Amflora provides a plausible reason for commercially cultivation of GM Crops in Europe. For improving the industrial application of potato, the amylose-less Amflora was genetically engineered to produce only amylopectin component in its starch. Amflora was developed by silencing the expression of the starch synthase enzyme using antisense strategy to eliminate the expression of amylose. Amylopectin is known to be the required starch component for industrial purpose because of its thickening properties, in the ...
We have developed an efficient method for transformation and regeneration of plants from carnatio... more We have developed an efficient method for transformation and regeneration of plants from carnation,Dianthus caryophyllus L. Whole leaves fromin vitro shoot cultures were mixed withAgrobacterium, cocultivated for 5 days and then plated on 2 µg/l chlorsulfuron (CS). Regenerated shoots and shoot clusters were divided into smaller sections and plated on 3 µg/l CS for selection to produce fully transformed shoots.
The major objective of our cucumber breeding program involves breeding for CMV disease resistance... more The major objective of our cucumber breeding program involves breeding for CMV disease resistance to reduce losses in quality and yield. Seven half diallel cross hybrids resistant to CMV were developed at the Vegetables Breeding Dept., Hort. Res. Inst., Egypt during March, 2006. Sixteen genetically different pure inbred lines of Cucumis sativus were used to develop the hybrids obtained from different sources and selected for their CMV disease-resistance trait. The inbred lines and one commercial (Beit alpha) cultivar as a ...
Berichte aus dem Julius Kühn-Institut, Nov 21, 2011
Larvae of the genus Agrotis (Lepidoptera: Noctuidae) are known to be severe soil pests on a wide ... more Larvae of the genus Agrotis (Lepidoptera: Noctuidae) are known to be severe soil pests on a wide range of field crops and vegetables in Europe, Asia and Africa. Agrotis spec. are highly susceptible for a broad number of baculoviruses and in the past, two Alphabaculoviruses (AgseNPV-A and AgseNPV-B) and one Betabaculovirus (AgseGV) were isolated from the common cutworm A. segetum. From larvae of the black cutworm A. ipsilon another
Corn borers are serious insect pests in many corn growing areas in Egypt and are responsible for ... more Corn borers are serious insect pests in many corn growing areas in Egypt and are responsible for significant yield losses. The ability of insect chitinases for attacking and digesting insect chitin raises the idea of using it for controlling insects such as corn borers. In this study, insect chitinases have been extracted from molting fluid as well as from the integument of Sesamia cretica (pink borer) and subjected to protein purification techniques, through three steps of precipitations 40, 60 and 80% of ammonium sulfate saturation. ...
Additional file 3: Figure S3. Digestion of pHT01 plasmid and the amplified kIspS fragment with Ba... more Additional file 3: Figure S3. Digestion of pHT01 plasmid and the amplified kIspS fragment with BamHI and XbaI to 11 generate pHT01-kIspS construct. Marker: 2 log DNA ladder (1.0â 10.0 kb) NEB catalogue #N04695, 12 Lane 1: pHT01 (7.9 kb) digested by BamHI and XbaI, Lane 2: amplified kIspS fragment (1.7 kb) 13 digested by BamHI and XbaI.
Additional file 2: Figure S2. Colony PCR results of the recombinant plasmid pET28b-kIspS-C term i... more Additional file 2: Figure S2. Colony PCR results of the recombinant plasmid pET28b-kIspS-C term in BL21 (DE3) cells. 7 Marker: 2 log DNA ladder (1.0â 10.0 kb) NEB catalogue #N3200, Lane 1, 2 & 3 are positive results for 8 the colony PCR of pET28b-kIspS-C terminal in BL21 (DE3) cells.
Apigenin is one of the most studied flavonoids and is widely distributed in the plant kingdom. Ap... more Apigenin is one of the most studied flavonoids and is widely distributed in the plant kingdom. Apigenin exerts important antioxidant, antibacterial, antifungal, antitumor activities, and anti-inflammatory effects in neurological or cardiovascular disease. Chalcone isomerase A (chiA) is an important enzyme of the flavonoid biosynthesis pathway. In order to enhance the apigenin production, the petunia chi A gene was transformed for Astragalus trigonus. Bialaphos survived plants were screened by PCR, dot blot hybridization and RT-PCR analysis. Also, jasmonic acid, salicylic acid, chitosan and yeast extract were tested to evaluate their capacity to work as elicitors for apigenin. Results showed that yeast extract was the best elicitor for induction of apigenin with an increase of 3.458 and 3.9 fold of the control for calli and cell suspension culture, respectively. Transformed cell suspension showed high apigenin content with a 20.17 fold increase compared to the control and 6.88 fold m...
3 Abstract: Influenza A viruses infects birds, animals and humans and it has the ability to mutat... more 3 Abstract: Influenza A viruses infects birds, animals and humans and it has the ability to mutate and develop mutant variants which may cause unexpected events. There is an urgent need for new medications to combat influenza pandemics. Using the genome sequences of the influenza A virus performed previously, we designed and performed a combinatorial exhaustive systematic methodology for optimal design of universal therapeutic short-interfering RNA (siRNA) molecules targeting all diverse of the virus strains. By using the bioinformatics tools, we designed siRNA targeting influenza A virus polymerase genes. The selection and validation of optimal siRNAs was also studied. The influenza A virus subtype H1N1 and H5N1 were propagated in MDCK cells. TCID and HA assays were performed and real time PCR for assessment of viral load was used. Time- 50 dependent and dosage-dependent usage were performed and showed high decrease in viral load.
Plant defensins are small cysteine-rich peptides which belong to a group of pathogenasis related ... more Plant defensins are small cysteine-rich peptides which belong to a group of pathogenasis related defense mechanism proteins. The proteins inhibit the growth of a broad range of microbes and are highly stable under extreme environmental stresses. Tomato cultivation is affected by fungal disease such as Fusarium wilt. In order to overcome fungal damages, transgenic tomato plants expressing the Medicago sativa defensin gene MsDef1 under the control of the CaMV 35S promoter were developed. The Fusarium-susceptible tomato (Lycobersicum esculentum Mill) cultivar CastleRock was used for transformation to acquire fungal resistance. Hypocotyl with a part of cotyledon (hypocotyledonary) for young tomato seedlings were used as an explant material and transformation was performed using the biolistic delivery system. Bombarded shoots were selected on regeneration medium supplemented with hygromycin and suitable concentrations of BA, zeatin ripozide and AgNO(3). Putative transgenic plantlets of T(0) were confirmed by PCR analysis using primers specific for the transgene and the transformation frequency obtained was 52.3%. Transformation and transcription of transgenes were confirmed in T(1) by PCR, Southern hybridizations, and reverse-transcription PCR (RT-PCR). The copy numbers of integrated transgene into tomato genome ranged between 1-3 copies. Greenhouse bioassay was performed on the transgenic T(1) and T(2) young seedlings and non-transgenic controls by challenging with a vigorous isolate of the fungal pathogen Fusarium oxysporum f. sp. Lycopersici. The level of fungal infectivity was determined using RT-PCR with tomatinase specific primers. Transgenic lines were more resistant to infection by fusarium than the control plants. These results indicated that overexpressing defensins in transgenic plants confer resistance to fungal pathogens.
Abstract: Tuesday the 25 th of January is an enshrined date in Egypt, it will be always remembere... more Abstract: Tuesday the 25 th of January is an enshrined date in Egypt, it will be always remembered as the date Egyptians decided to fight for their freedom. Young Egyptians took to the streets and main squares. Their gathering ignited the revolution as three days later millions of protesters further gathered to demand changing the regime. They asked for “freedom and human dignity”. However, their demands rose to the resigning of Mubarak after unnecessary use of force by the police. Eighteen days later Mubarak stepped down ...
In Africa, six million children die from malnutrition before their fifth birthday each year (Worl... more In Africa, six million children die from malnutrition before their fifth birthday each year (World Bank statistics). More than 800 million people go to bed hungry, 300 million of them are children. “Simply because my people are hungry, that is no justification to give them poison.” Small words that were spoken by President Levy Mwanawasa of Zambia, who refused to accept milled grains from the United States during the worst food shortage crisis that Africa ever faced in 2002. Amazingly, African leaders of Zambia, Zimbabwe, Malawi and ...
In Africa, six million children die from malnutrition before their fifth birthday each year (Worl... more In Africa, six million children die from malnutrition before their fifth birthday each year (World Bank statistics). More than 800 million people go to bed hungry, 300 million of them are children. “Simply because my people are hungry, that is no justification to give them poison.” Small words that were spoken by President Levy Mwanawasa of Zambia, who refused to accept milled grains from the United States during the worst food shortage crisis that Africa ever faced in 2002. Amazingly, African leaders of Zambia, Zimbabwe, Malawi and ...
Abstract: Amflora provides a plausible reason for commercially cultivation of GM Crops in Europe.... more Abstract: Amflora provides a plausible reason for commercially cultivation of GM Crops in Europe. For improving the industrial application of potato, the amylose-less Amflora was genetically engineered to produce only amylopectin component in its starch. Amflora was developed by silencing the expression of the starch synthase enzyme using antisense strategy to eliminate the expression of amylose. Amylopectin is known to be the required starch component for industrial purpose because of its thickening properties, in the ...
We have developed an efficient method for transformation and regeneration of plants from carnatio... more We have developed an efficient method for transformation and regeneration of plants from carnation,Dianthus caryophyllus L. Whole leaves fromin vitro shoot cultures were mixed withAgrobacterium, cocultivated for 5 days and then plated on 2 µg/l chlorsulfuron (CS). Regenerated shoots and shoot clusters were divided into smaller sections and plated on 3 µg/l CS for selection to produce fully transformed shoots.
The major objective of our cucumber breeding program involves breeding for CMV disease resistance... more The major objective of our cucumber breeding program involves breeding for CMV disease resistance to reduce losses in quality and yield. Seven half diallel cross hybrids resistant to CMV were developed at the Vegetables Breeding Dept., Hort. Res. Inst., Egypt during March, 2006. Sixteen genetically different pure inbred lines of Cucumis sativus were used to develop the hybrids obtained from different sources and selected for their CMV disease-resistance trait. The inbred lines and one commercial (Beit alpha) cultivar as a ...
Berichte aus dem Julius Kühn-Institut, Nov 21, 2011
Larvae of the genus Agrotis (Lepidoptera: Noctuidae) are known to be severe soil pests on a wide ... more Larvae of the genus Agrotis (Lepidoptera: Noctuidae) are known to be severe soil pests on a wide range of field crops and vegetables in Europe, Asia and Africa. Agrotis spec. are highly susceptible for a broad number of baculoviruses and in the past, two Alphabaculoviruses (AgseNPV-A and AgseNPV-B) and one Betabaculovirus (AgseGV) were isolated from the common cutworm A. segetum. From larvae of the black cutworm A. ipsilon another
Corn borers are serious insect pests in many corn growing areas in Egypt and are responsible for ... more Corn borers are serious insect pests in many corn growing areas in Egypt and are responsible for significant yield losses. The ability of insect chitinases for attacking and digesting insect chitin raises the idea of using it for controlling insects such as corn borers. In this study, insect chitinases have been extracted from molting fluid as well as from the integument of Sesamia cretica (pink borer) and subjected to protein purification techniques, through three steps of precipitations 40, 60 and 80% of ammonium sulfate saturation. ...
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