In order to investigate the functions of the gene PsG6PDH and the mechanisms underlying freezing tolerance of Populus suaveolens, the recombinant expression vector pET-G （pET30a-G6PDH）, which contained full encoding...In order to investigate the functions of the gene PsG6PDH and the mechanisms underlying freezing tolerance of Populus suaveolens, the recombinant expression vector pET-G （pET30a-G6PDH）, which contained full encoding region of PsG6PDH gene, was established. The recombinant was identified by lawn-PCR and double enzyme digestion and then transformed into expression host XA90 and induced by isopropyl-a-D-thiogalactoside （IPTG） to express 100 kD polypeptide of G6PDH fusion protein. The results showed that the expressed amount of the fusion protein culminated after 1 mmol·L^-1 IPTG treatment for 4h and that pET-G product was predominately soluble and not extra-cellular secreting.展开更多
Glucose 6 phosphate dehydrogenase(G6PD)is a key and rate limiting enzyme in the pentose phosphate pathway(PPP).The physiological significance of enzyme is providing reduced energy to specific cells like erythrocyte by...Glucose 6 phosphate dehydrogenase(G6PD)is a key and rate limiting enzyme in the pentose phosphate pathway(PPP).The physiological significance of enzyme is providing reduced energy to specific cells like erythrocyte by maintaining co-enzyme nicotinamide adenine dinucleotide phosphate(NADPH).There are preponderance research findings that demonstrate the enzyme(G6PD)role in the energy balance,and it is associated with bloodrelated diseases and disorders,primarily the anemia resulted from G6PD deficiency.The Xlinked genetic deficiency of G6PD and associated non-immune hemolytic anemia have been studied widely across the globe.Recent advancement in biology,more precisely neuroscience has revealed that G6PD is centrally involved in many neurological and neurodegenerative disorders.The neuroprotective role of the enzyme(G6PD)has also been established,as well as the potential of G6PD in oxidative damage and the Reactive Oxygen Species(ROS)produced in cerebral ischemia.Though G6PD deficiency remains a global health issue,however,a paradigm shift in research focusing the potential of the enzyme in neurological and neurodegenerative disorders will surely open a new avenue in diagnostics and enzyme therapeutics.Here,in this study,more emphasis was made on exploring the role of G6PD in neurological and inflammatory disorders as well as non-immune hemolytic anemia,thus providing diagnostic and therapeutic opportunities.展开更多
Objective To prepare a kit for screening individuals with glucose 6 phosphate dehydrogenase (G6PD) defect. The kit is easy to use and to get the fast as well as reliable results. Especially it is suitable for the a...Objective To prepare a kit for screening individuals with glucose 6 phosphate dehydrogenase (G6PD) defect. The kit is easy to use and to get the fast as well as reliable results. Especially it is suitable for the anti malaria spots usually located in the remote countryside where no electricity is available. Methods The double filter paper method and other 2 techniques, the quantitative method and the single filter paper method, were used to determine G6PD activity in 70 samples of human erythrocytes. It was found that the results of the double filter paper method and those of the single filter paper method in the first 8 hours after the drying of the blood soaked filter paper were consistent with those of the quantitative method. When a piece of blood soaked paper is left under room temperature more than 24 hours, G6PD in the erythrocytes deteriorated spontaneously and consequently the number of positive cases increased along with the elapse of time.Results Satisfactory results were achieved when the kit was used to screen cases of G6PD defect from 151 farmers who were receiving anti mararia therapy. The kit was made according to a technique named “double filter paper” method.Conclusions These findings suggest that the double filter paper method can reveal the level of G6PD activity and the results are rapidly obtained when the method is used on the anti malaria spot.展开更多
文摘In order to investigate the functions of the gene PsG6PDH and the mechanisms underlying freezing tolerance of Populus suaveolens, the recombinant expression vector pET-G （pET30a-G6PDH）, which contained full encoding region of PsG6PDH gene, was established. The recombinant was identified by lawn-PCR and double enzyme digestion and then transformed into expression host XA90 and induced by isopropyl-a-D-thiogalactoside （IPTG） to express 100 kD polypeptide of G6PDH fusion protein. The results showed that the expressed amount of the fusion protein culminated after 1 mmol·L^-1 IPTG treatment for 4h and that pET-G product was predominately soluble and not extra-cellular secreting.
文摘Glucose 6 phosphate dehydrogenase(G6PD)is a key and rate limiting enzyme in the pentose phosphate pathway(PPP).The physiological significance of enzyme is providing reduced energy to specific cells like erythrocyte by maintaining co-enzyme nicotinamide adenine dinucleotide phosphate(NADPH).There are preponderance research findings that demonstrate the enzyme(G6PD)role in the energy balance,and it is associated with bloodrelated diseases and disorders,primarily the anemia resulted from G6PD deficiency.The Xlinked genetic deficiency of G6PD and associated non-immune hemolytic anemia have been studied widely across the globe.Recent advancement in biology,more precisely neuroscience has revealed that G6PD is centrally involved in many neurological and neurodegenerative disorders.The neuroprotective role of the enzyme(G6PD)has also been established,as well as the potential of G6PD in oxidative damage and the Reactive Oxygen Species(ROS)produced in cerebral ischemia.Though G6PD deficiency remains a global health issue,however,a paradigm shift in research focusing the potential of the enzyme in neurological and neurodegenerative disorders will surely open a new avenue in diagnostics and enzyme therapeutics.Here,in this study,more emphasis was made on exploring the role of G6PD in neurological and inflammatory disorders as well as non-immune hemolytic anemia,thus providing diagnostic and therapeutic opportunities.
文摘Objective To prepare a kit for screening individuals with glucose 6 phosphate dehydrogenase (G6PD) defect. The kit is easy to use and to get the fast as well as reliable results. Especially it is suitable for the anti malaria spots usually located in the remote countryside where no electricity is available. Methods The double filter paper method and other 2 techniques, the quantitative method and the single filter paper method, were used to determine G6PD activity in 70 samples of human erythrocytes. It was found that the results of the double filter paper method and those of the single filter paper method in the first 8 hours after the drying of the blood soaked filter paper were consistent with those of the quantitative method. When a piece of blood soaked paper is left under room temperature more than 24 hours, G6PD in the erythrocytes deteriorated spontaneously and consequently the number of positive cases increased along with the elapse of time.Results Satisfactory results were achieved when the kit was used to screen cases of G6PD defect from 151 farmers who were receiving anti mararia therapy. The kit was made according to a technique named “double filter paper” method.Conclusions These findings suggest that the double filter paper method can reveal the level of G6PD activity and the results are rapidly obtained when the method is used on the anti malaria spot.
