Neisseria meningitides is a Gram-negative bacterium which is an important causative agent of septicemia and meningitis.Numerous pathogenic bacteria contain luxS,which is required for autoinducer-2 production.Neisseria...Neisseria meningitides is a Gram-negative bacterium which is an important causative agent of septicemia and meningitis.Numerous pathogenic bacteria contain luxS,which is required for autoinducer-2 production.Neisseria meningitides contains a functional copy of luxS that is necessary for full meningococcal virulence.Neisseria meningitides DNA was extracted and its luxS gene was amplified by nested PCR.PCR product was purified and cloned in to pQE-30 expression vector.Recombinant plasmid was transformed and mass cultured.luxS was expressed in E.coli and confirmed by western blot analysis.In this study,N.meningitides luxS was amplified, cloned and expressed successfully.The sequencing of PCR product confirmed that amplified gene was luxS. Gene was expressed and observed in SDS-PAGE.Protein was reacted by his tag monoclonal antibody through western blot analysis.展开更多
The Caspian bent-toed gecko, Tenuidactylus caspius, belonging to the family Gekkonidae, is widely distributed across the northern half of Iran, especially along the southern coastal region of the Caspian Sea. It is re...The Caspian bent-toed gecko, Tenuidactylus caspius, belonging to the family Gekkonidae, is widely distributed across the northern half of Iran, especially along the southern coastal region of the Caspian Sea. It is regarded as a commensal species throughout its entire distribution. We investigated genetic variation and relationships among different populations of this species in Iran. Fragments of the mitochondrial cyt b（547 bp） and ND4（831 bp） genes were sequenced and analyzed in 64 and 28 specimens, respectively, from 21 geographically distant localities. Cyrtopodion scabrum was used as the outgroup taxon. The data showed no significant genetic variation within the populations of T. caspius in Iran. Nevertheless, populations of Khorasan and Semnan（especially Shahrood） in northeastern Iran showed greater divergence（p-distance = 2.1%） from other Iranian populations. The low genetic variation and homogeneous structure among populations of T. caspius on either side of the Elburz Mountains suggests that this species most likely has achieved its current distribution recently and as a result of anthropogenic activities.展开更多
Objective:To isolate and characterize the saponin from Persian Gulf brittle star(Ophiocoma erinaceus)and to evaluate its hemolytic and cytotoxic potential. Methods:In an attempt to prepare saponin from brittle star,co...Objective:To isolate and characterize the saponin from Persian Gulf brittle star(Ophiocoma erinaceus)and to evaluate its hemolytic and cytotoxic potential. Methods:In an attempt to prepare saponin from brittle star,collected samples were minced and extracted with ethanol,dichloromethane,n-buthanol.Then,concentrated n-butanol extract were loaded on HP-20 resin and washed with dionized water,80%ethanol and 100%ethanol respectively.Subsequently,detection of saponin was performed by foaming property,fourier transform infrared spectroscopy and hemolytic analysis on thin layer chromatography.The cytotoxic activity on HeLa cells was evaluated through 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazoliumbromide(MTT)assay and under invert microscopy.Results:The existence of saponin in Ophiocoma erinaceus were approved by phytochemical method.The presence of C-H bond,C-O-C and OH in fourier transform infrared spectrum of fraction 80%ethanol is characteristic feature in the many of saponin compounds.Hemolytic assay revealed HD_(50) value was 500μg/mL.MTT assay exhibited that saponin extracted in IC_(50) value of 25μg/mL inducsd potent cytotoxic activity against HeLa cells in 24 h and 12.5μg/mL in 48 h,meanwhile in lower concentration did not have considerable effect against HeLa cells.Conclusions:These findings showed that only 80%ethanol fraction Persian Gulf brittle star contained saponin like compounds with hemolytic activity which can be detected simply by phytochemical that can be appreciable for future anticancer research.展开更多
基金supported by Vice Chancellor of Research of Shahid Beheshti University M.C.and was done in Cellular and Molecular Biology Research Center
文摘Neisseria meningitides is a Gram-negative bacterium which is an important causative agent of septicemia and meningitis.Numerous pathogenic bacteria contain luxS,which is required for autoinducer-2 production.Neisseria meningitides contains a functional copy of luxS that is necessary for full meningococcal virulence.Neisseria meningitides DNA was extracted and its luxS gene was amplified by nested PCR.PCR product was purified and cloned in to pQE-30 expression vector.Recombinant plasmid was transformed and mass cultured.luxS was expressed in E.coli and confirmed by western blot analysis.In this study,N.meningitides luxS was amplified, cloned and expressed successfully.The sequencing of PCR product confirmed that amplified gene was luxS. Gene was expressed and observed in SDS-PAGE.Protein was reacted by his tag monoclonal antibody through western blot analysis.
基金supported by the Iran National Scientific Foundation(INSF) under proposal number of 89001493
文摘The Caspian bent-toed gecko, Tenuidactylus caspius, belonging to the family Gekkonidae, is widely distributed across the northern half of Iran, especially along the southern coastal region of the Caspian Sea. It is regarded as a commensal species throughout its entire distribution. We investigated genetic variation and relationships among different populations of this species in Iran. Fragments of the mitochondrial cyt b（547 bp） and ND4（831 bp） genes were sequenced and analyzed in 64 and 28 specimens, respectively, from 21 geographically distant localities. Cyrtopodion scabrum was used as the outgroup taxon. The data showed no significant genetic variation within the populations of T. caspius in Iran. Nevertheless, populations of Khorasan and Semnan（especially Shahrood） in northeastern Iran showed greater divergence（p-distance = 2.1%） from other Iranian populations. The low genetic variation and homogeneous structure among populations of T. caspius on either side of the Elburz Mountains suggests that this species most likely has achieved its current distribution recently and as a result of anthropogenic activities.
文摘Objective:To isolate and characterize the saponin from Persian Gulf brittle star(Ophiocoma erinaceus)and to evaluate its hemolytic and cytotoxic potential. Methods:In an attempt to prepare saponin from brittle star,collected samples were minced and extracted with ethanol,dichloromethane,n-buthanol.Then,concentrated n-butanol extract were loaded on HP-20 resin and washed with dionized water,80%ethanol and 100%ethanol respectively.Subsequently,detection of saponin was performed by foaming property,fourier transform infrared spectroscopy and hemolytic analysis on thin layer chromatography.The cytotoxic activity on HeLa cells was evaluated through 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazoliumbromide(MTT)assay and under invert microscopy.Results:The existence of saponin in Ophiocoma erinaceus were approved by phytochemical method.The presence of C-H bond,C-O-C and OH in fourier transform infrared spectrum of fraction 80%ethanol is characteristic feature in the many of saponin compounds.Hemolytic assay revealed HD_(50) value was 500μg/mL.MTT assay exhibited that saponin extracted in IC_(50) value of 25μg/mL inducsd potent cytotoxic activity against HeLa cells in 24 h and 12.5μg/mL in 48 h,meanwhile in lower concentration did not have considerable effect against HeLa cells.Conclusions:These findings showed that only 80%ethanol fraction Persian Gulf brittle star contained saponin like compounds with hemolytic activity which can be detected simply by phytochemical that can be appreciable for future anticancer research.
