R Hernández-Escamilla, J M Espinosa-Lara, J D Quintana-Hau, S Uribe-Luna, P Loyola-Abitia, J C Santiago-Hernández, R Maldonado-Rodríguez
The aim of this work was to search if the rat DNA polymerase beta can substitute the capability of DNA polymerase I to repair damage caused by the UV light in Escherichia coli. The oriC origin of replication from p beta 5 was replaced by the rep origin from pSC101 and named p beta 6. The presence of pol beta in the new construct was verified by PCR. E. coli polA-1 (WP6) was transformed with p beta 6. A protein with size similar to DNA Pol beta (40 kDa) was shown in the cell free extracts carrying pbeta5. In WP6/p beta 6 cell free extracts a slightly smaller protein was observed instead of the 40 kDa. DNA Pol beta was revealed by western analysis, with polyclonal antibodies, in strains with p beta 5. Yet, it was not detected in the western from WP6/p beta 6. A moderate change in UV resistance was observed in strains carrying p beta 5. However, in polAl carrying p beta 6 (WP6/p beta 6), irradiated with 60-90 J/m2 of UV light, the viability was increased by more than four orders of magnitude, when compared with the polA1 (WP6) strain, reaching approximately the same UV resistance as the strains with DNA polymerase I. The results suggests that probably Pol beta is rapidly degraded in the cell free extracts from WP6/p beta 6 and, it repairs the lethal effect of the UV light in E. coli.
{"title":"Rat DNA polymerase beta substitutes the repairing activity of DNA polymerase I in the lethal effect of UV light.","authors":"R Hernández-Escamilla, J M Espinosa-Lara, J D Quintana-Hau, S Uribe-Luna, P Loyola-Abitia, J C Santiago-Hernández, R Maldonado-Rodríguez","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>The aim of this work was to search if the rat DNA polymerase beta can substitute the capability of DNA polymerase I to repair damage caused by the UV light in Escherichia coli. The oriC origin of replication from p beta 5 was replaced by the rep origin from pSC101 and named p beta 6. The presence of pol beta in the new construct was verified by PCR. E. coli polA-1 (WP6) was transformed with p beta 6. A protein with size similar to DNA Pol beta (40 kDa) was shown in the cell free extracts carrying pbeta5. In WP6/p beta 6 cell free extracts a slightly smaller protein was observed instead of the 40 kDa. DNA Pol beta was revealed by western analysis, with polyclonal antibodies, in strains with p beta 5. Yet, it was not detected in the western from WP6/p beta 6. A moderate change in UV resistance was observed in strains carrying p beta 5. However, in polAl carrying p beta 6 (WP6/p beta 6), irradiated with 60-90 J/m2 of UV light, the viability was increased by more than four orders of magnitude, when compared with the polA1 (WP6) strain, reaching approximately the same UV resistance as the strains with DNA polymerase I. The results suggests that probably Pol beta is rapidly degraded in the cell free extracts from WP6/p beta 6 and, it repairs the lethal effect of the UV light in E. coli.</p>","PeriodicalId":21464,"journal":{"name":"Revista latinoamericana de microbiologia","volume":"44 2","pages":"58-64"},"PeriodicalIF":0.0,"publicationDate":"2002-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"26329141","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Gloria Luz Paniagua, Eric Monroy, Erasmo Negrete, Sergio Vaca
Eighty Candida albicans strains, isolated from throat of patients at the Universitary Clinic of the Faculty of Superior Studies Iztacala of the National Autonomous University of Mexico, were analyzed. They were identified by microscopic and colony morphologies, germ tube test, and by auxanogram and zimogram. Minimal inhibitory concentrations (MIC) of 5-fluorocytosine, miconazole and amphotericin B were determined by microtiter broth dilution. MIC frequency distribution of 5-fluorocytosine showed a single peak (0.25-8.0 microg/ml), with 65% susceptible strains (MIC < or = 1.0 < or =g/ml) and 35% intermediate susceptible strains (MIC = 1.1-8 microg/ml). MIC frequency distribution of miconazole was threemodal with 6.25% susceptible (MIC = 1.562 microg/ml), 48.75% intermediate susceptible (MIC = 3.125-12.5 microg/ml), and 45% resistant (MIC = 25-50 microg/ml) strains. All strains were susceptible to amphotericin B (MIC= 0.0156-0.125 microg/m). These results shows that amphotericin B was the more active antimycotic, followed by 5-fluorocytosine, against the strains analyzed, and that miconazole was the less effective one.
{"title":"Susceptibility to 5-fluorocytosine, miconazole and amphotericin B of Candida albicans strains isolated from the throat of non-AIDS patients.","authors":"Gloria Luz Paniagua, Eric Monroy, Erasmo Negrete, Sergio Vaca","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>Eighty Candida albicans strains, isolated from throat of patients at the Universitary Clinic of the Faculty of Superior Studies Iztacala of the National Autonomous University of Mexico, were analyzed. They were identified by microscopic and colony morphologies, germ tube test, and by auxanogram and zimogram. Minimal inhibitory concentrations (MIC) of 5-fluorocytosine, miconazole and amphotericin B were determined by microtiter broth dilution. MIC frequency distribution of 5-fluorocytosine showed a single peak (0.25-8.0 microg/ml), with 65% susceptible strains (MIC < or = 1.0 < or =g/ml) and 35% intermediate susceptible strains (MIC = 1.1-8 microg/ml). MIC frequency distribution of miconazole was threemodal with 6.25% susceptible (MIC = 1.562 microg/ml), 48.75% intermediate susceptible (MIC = 3.125-12.5 microg/ml), and 45% resistant (MIC = 25-50 microg/ml) strains. All strains were susceptible to amphotericin B (MIC= 0.0156-0.125 microg/m). These results shows that amphotericin B was the more active antimycotic, followed by 5-fluorocytosine, against the strains analyzed, and that miconazole was the less effective one.</p>","PeriodicalId":21464,"journal":{"name":"Revista latinoamericana de microbiologia","volume":"44 2","pages":"65-8"},"PeriodicalIF":0.0,"publicationDate":"2002-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"26329142","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
M P Crisóstomo-Vázquez, M P Cervantes-Cervantes, E Jiménez-Cardoso, J L Muñoz-Sánchez
Entamoeba histolytica is a parasite which causes health problems and there has been many approaches to know what is the factor causing its pathogenicity. In the present work, we assayed if the production of free radicals by the amoeba, has a relationship with the proteases activity. When we test the DMSO action (free radicals quenching activity) the specific activity of the proteases complex of the parasite were affected also. At 33.3% (V/V) concentration of DMSO it was present a maximal decrease of the initial activity (about 46% decrease), for to a higher concentrations existing a trend to recuperate the original activity, suggesting that the free radicals are an important factor for the hydrolysis grade of the protein substrate. All the differences except those between 46.7 and 66.6%, were significantly different compared with the control without any addition. The effects of Probucol and Probucol plus DMSO, compared to those caused by Metronidazol (MZ). We can observe that the quenchers caused a decrease on proteases activity similar to that of MZ (which is an antiparasite drug) and it was of c.a. 58% of activity decrease. These data suggest that the action of both, free radicals and proteases complex of Entamoeba histolytica, can account for the pathogenicity of the parasite.
{"title":"Relationship between free radicals produced by entamoeba histolytica and its proteases complex activity.","authors":"M P Crisóstomo-Vázquez, M P Cervantes-Cervantes, E Jiménez-Cardoso, J L Muñoz-Sánchez","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>Entamoeba histolytica is a parasite which causes health problems and there has been many approaches to know what is the factor causing its pathogenicity. In the present work, we assayed if the production of free radicals by the amoeba, has a relationship with the proteases activity. When we test the DMSO action (free radicals quenching activity) the specific activity of the proteases complex of the parasite were affected also. At 33.3% (V/V) concentration of DMSO it was present a maximal decrease of the initial activity (about 46% decrease), for to a higher concentrations existing a trend to recuperate the original activity, suggesting that the free radicals are an important factor for the hydrolysis grade of the protein substrate. All the differences except those between 46.7 and 66.6%, were significantly different compared with the control without any addition. The effects of Probucol and Probucol plus DMSO, compared to those caused by Metronidazol (MZ). We can observe that the quenchers caused a decrease on proteases activity similar to that of MZ (which is an antiparasite drug) and it was of c.a. 58% of activity decrease. These data suggest that the action of both, free radicals and proteases complex of Entamoeba histolytica, can account for the pathogenicity of the parasite.</p>","PeriodicalId":21464,"journal":{"name":"Revista latinoamericana de microbiologia","volume":"44 2","pages":"79-82"},"PeriodicalIF":0.0,"publicationDate":"2002-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"26329145","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
José Antonio Rivera-Tapia, María Lilia Cedillo-Ramírez, Constantino Gil Juárez
Mycoplasmas are a bacterial group that is classified in the Mollicute class which includes Mycoplasmas, Spiroplasmas and Acholeplasmas. One hundred and seventy six species have been described in this group. Mycoplasmas are the smallest self living prokaryotes, they do not have a bacterial wall, their genomic size ranges from 577 to 2220 bpk, they are nutritional exigent so it is hard to culture them, but the development of molecular biology techniques has let us detect more mycoplasmas in different hosts. Mycoplasmas have been associated to acute and chronic diseases mainly in animals and humans while spiroplasmas have been found in arthropods, plants and flowers producing or not damage. Some recent studies have shown the role of some structural components of Mycoplasmas in pathogenesis, such as cytoskeleton proteins and adhesins, and the influence of some genetic characteristics on the development of an infectious disease.
{"title":"Some biological features of mollicutes.","authors":"José Antonio Rivera-Tapia, María Lilia Cedillo-Ramírez, Constantino Gil Juárez","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>Mycoplasmas are a bacterial group that is classified in the Mollicute class which includes Mycoplasmas, Spiroplasmas and Acholeplasmas. One hundred and seventy six species have been described in this group. Mycoplasmas are the smallest self living prokaryotes, they do not have a bacterial wall, their genomic size ranges from 577 to 2220 bpk, they are nutritional exigent so it is hard to culture them, but the development of molecular biology techniques has let us detect more mycoplasmas in different hosts. Mycoplasmas have been associated to acute and chronic diseases mainly in animals and humans while spiroplasmas have been found in arthropods, plants and flowers producing or not damage. Some recent studies have shown the role of some structural components of Mycoplasmas in pathogenesis, such as cytoskeleton proteins and adhesins, and the influence of some genetic characteristics on the development of an infectious disease.</p>","PeriodicalId":21464,"journal":{"name":"Revista latinoamericana de microbiologia","volume":"44 2","pages":"53-7"},"PeriodicalIF":0.0,"publicationDate":"2002-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"26387429","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
J A Salinas-Melédez, R Avalos-Ramírez, V Riojas-Valdez, J Kawas-Garza, H Fimbres-Durazo, G Hernández-Vidal
The serological prevalence of Q fever in Mexico is unknown. A serological survey for Coxiella burnetii was undertaken on a randomly selected population of dairy cattle, beef cattle, goats and sheep flocks. Serological examination of animal sera for antibodies against Coxiella burnetii was carried out by the ELISA technique. The 28% of the dairy cattle and 10% of beef cattle examinated were antibody positive. Sera from goats and sheep also had antibodies against this rickettsia, 35% and 40% respectively.
{"title":"Serologic survey in animals of 'Q' fever in Nuevo Leon.","authors":"J A Salinas-Melédez, R Avalos-Ramírez, V Riojas-Valdez, J Kawas-Garza, H Fimbres-Durazo, G Hernández-Vidal","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>The serological prevalence of Q fever in Mexico is unknown. A serological survey for Coxiella burnetii was undertaken on a randomly selected population of dairy cattle, beef cattle, goats and sheep flocks. Serological examination of animal sera for antibodies against Coxiella burnetii was carried out by the ELISA technique. The 28% of the dairy cattle and 10% of beef cattle examinated were antibody positive. Sera from goats and sheep also had antibodies against this rickettsia, 35% and 40% respectively.</p>","PeriodicalId":21464,"journal":{"name":"Revista latinoamericana de microbiologia","volume":"44 2","pages":"75-8"},"PeriodicalIF":0.0,"publicationDate":"2002-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"26329146","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The analysis of behavior of man in the field of biology is carried out through bioethics, considered the science of the survival. In the microbiology, there are numerous discoveries related with pathogenic microorganisms, including those that can be used as weapons in a biological war or in an attack considered bioterrorism. The scientist involved in microbiology can participate with his knowledge in the development and improvement of bioweapons, however from the point of view of bioethics it is not acceptable that he works in an investigation related with these topics, because the defense research can evolve in offensive one. The war is an antisurvival activity, therefore it is not acceptable. In the same way, the biological weapons composed with virus, fungi or alive bacteria, or with toxins from them, neither they are morally accepted. After the terrorist attacks with anthrax in the United States in 2001, the world scientific community in the field of microbiology should show against the use of the microorganisms like bioweapons, at the time of promoting the idea that the responsible use for the microorganisms is a moral imperative for all microbiologists around the world, since the biological weapons are a threat for the human life.
{"title":"Bioethics, bioweapons and the microbiologist.","authors":"Fernando Anaya-Velázquez","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>The analysis of behavior of man in the field of biology is carried out through bioethics, considered the science of the survival. In the microbiology, there are numerous discoveries related with pathogenic microorganisms, including those that can be used as weapons in a biological war or in an attack considered bioterrorism. The scientist involved in microbiology can participate with his knowledge in the development and improvement of bioweapons, however from the point of view of bioethics it is not acceptable that he works in an investigation related with these topics, because the defense research can evolve in offensive one. The war is an antisurvival activity, therefore it is not acceptable. In the same way, the biological weapons composed with virus, fungi or alive bacteria, or with toxins from them, neither they are morally accepted. After the terrorist attacks with anthrax in the United States in 2001, the world scientific community in the field of microbiology should show against the use of the microorganisms like bioweapons, at the time of promoting the idea that the responsible use for the microorganisms is a moral imperative for all microbiologists around the world, since the biological weapons are a threat for the human life.</p>","PeriodicalId":21464,"journal":{"name":"Revista latinoamericana de microbiologia","volume":"44 1","pages":"38-45"},"PeriodicalIF":0.0,"publicationDate":"2002-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"26327328","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Alejandro Alarcón, Frederick T Davies, Johnatan N Egilla, Theodore C Fox, Arturo A Estrada-Luna, Ronald Ferrera-Cerrato
Arbuscular mycorrhizal fungi (AMF) are able to increase root enzymatic activity of acid and alkaline phosphatases. However, the role of AMF on phosphatase activity has not been reported in papaya (Carica papaya L.), which is frequently established at places with soil phosphorus (P) deficiencies. The goals of this research were to determine the effect of Glomus claroideum (Gc), and plant growth promoting rhizobacterium Azospirillum brasilense strain VS7 [Ab]) on root phosphatase activity and seedling growth of Carica papaya L. cv. Red Maradol under low P conditions. There were four treatments-colonization with: 1) Gc, 2) Ab, 3) Gc+Ab, and 4) non-inoculated seedlings. Plants were established in a coarse sand:sandy loam substrate under P-limitation (11 microg P ml(-1)), supplied with a modified Long Ashton Nutrient Solution. Seedling growth was severely reduced by low P. Gc+Ab inoculated plants had greater total dry matter and leaf area than non-colonized plants. Gc-inoculated plants had greater leaf area than non-colonized plants. Treatments did not differ in leaf area ratio, specific leaf area and, total chlorophyll content. There was a non-significant effect on stem relative growth rate with Gc and Gc+Ab plants. Mycorrhizal colonization enhanced the bacterial population 3.4-fold in the Gc+Ab treatment compared with the population quantified in Ab treatment. Soluble and extractable root acid phosphatase activity (RAPA) was higher in Gc inoculated plants. We discussed on the possible relation among both inoculated microorganisms and also with the P-limitation which plants were established.
丛枝菌根真菌(AMF)具有提高根内酸性和碱性磷酸酶活性的作用。然而,AMF对番木瓜(Carica papaya L.)磷酸酶活性的作用尚未报道,这通常是在土壤磷(P)缺乏的地方建立的。本研究旨在研究球囊菌(Glomus claroideum, Gc)和促生根瘤菌巴西氮螺旋菌VS7 [Ab]对番木瓜根系磷酸酶活性和幼苗生长的影响。低磷条件下的红马拉多尔。四种处理:1)Gc、2)Ab、3)Gc+Ab和4)未接种的幼苗定殖。植物生长在磷限制(11 μ g P ml(-1))的粗砂:砂壤土基质中,并提供改良的Long Ashton营养液。低P. Gc+Ab接种植株的总干物质和叶面积均大于非定殖植株。gc接种植株的叶面积大于非定殖植株。在叶面积比、比叶面积和总叶绿素含量方面,处理间无显著差异。Gc和Gc+Ab植株对茎相对生长率影响不显著。菌根定植使Gc+Ab处理的细菌数量比Ab处理的数量增加3.4倍。Gc接种植株的可溶性和可提取根酸性磷酸酶(RAPA)活性较高。我们讨论了接种微生物之间的可能关系以及植物所建立的磷限制。
{"title":"Short term effects of Glomus claroideum and Azospirillum brasilense on growth and root acid phosphatase activity of Carica papaya L. under phosphorus stress.","authors":"Alejandro Alarcón, Frederick T Davies, Johnatan N Egilla, Theodore C Fox, Arturo A Estrada-Luna, Ronald Ferrera-Cerrato","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>Arbuscular mycorrhizal fungi (AMF) are able to increase root enzymatic activity of acid and alkaline phosphatases. However, the role of AMF on phosphatase activity has not been reported in papaya (Carica papaya L.), which is frequently established at places with soil phosphorus (P) deficiencies. The goals of this research were to determine the effect of Glomus claroideum (Gc), and plant growth promoting rhizobacterium Azospirillum brasilense strain VS7 [Ab]) on root phosphatase activity and seedling growth of Carica papaya L. cv. Red Maradol under low P conditions. There were four treatments-colonization with: 1) Gc, 2) Ab, 3) Gc+Ab, and 4) non-inoculated seedlings. Plants were established in a coarse sand:sandy loam substrate under P-limitation (11 microg P ml(-1)), supplied with a modified Long Ashton Nutrient Solution. Seedling growth was severely reduced by low P. Gc+Ab inoculated plants had greater total dry matter and leaf area than non-colonized plants. Gc-inoculated plants had greater leaf area than non-colonized plants. Treatments did not differ in leaf area ratio, specific leaf area and, total chlorophyll content. There was a non-significant effect on stem relative growth rate with Gc and Gc+Ab plants. Mycorrhizal colonization enhanced the bacterial population 3.4-fold in the Gc+Ab treatment compared with the population quantified in Ab treatment. Soluble and extractable root acid phosphatase activity (RAPA) was higher in Gc inoculated plants. We discussed on the possible relation among both inoculated microorganisms and also with the P-limitation which plants were established.</p>","PeriodicalId":21464,"journal":{"name":"Revista latinoamericana de microbiologia","volume":"44 1","pages":"31-7"},"PeriodicalIF":0.0,"publicationDate":"2002-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"26326845","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Yeosvany Cabrera Artiles, Duniesky Martínez García, Enrique R Pérez Cruz, Gabriel J Márquez Perera, Manuel Luis Feble
The Thermus aquaticus DNA Polymerase I (Taq Pol I) gene was cloned into the pOSEX4 plasmid under the osmo-inducible promoter proU and subsequently expressed into the Escherichia coli MKH13 strain. The suitability of the enzyme in polymerase assays was determined in standard 35S dATP incorporation tests and by PCR. The Taq Pol I expression in this system, which is under the control of the osmotic pressure in the growth medium, was analyzed in different media and in different sodium chloride concentrations. A study of the osmolarity effects in the growth of the strain and in Taq Pol I expression shows that an increase in sodium chloride concentration limits the growth. At 0.25 M of NaCl maximum activity was observed; at higher values of osmolarity, we found an unexpected decline of activity. This is the first report of using the pOSEX vector for the expression of an heterologous protein and it is very advantageous to make a regulated, non toxic, simple and cost-effective manner of induction in a biotechnology process using just NaCl or other non-permeable osmolyte.
在渗透诱导启动子proU的作用下,将水热菌DNA聚合酶I (Taq Pol I)基因克隆到pOSEX4质粒中,并在大肠杆菌MKH13中表达。该酶在聚合酶试验中的适用性通过标准35S dATP掺入试验和PCR确定。在生长介质渗透压控制下,分析了该体系在不同培养基和不同氯化钠浓度下Taq Pol I的表达情况。渗透压对菌株生长和Taq Pol I表达的影响表明,氯化钠浓度的增加限制了菌株的生长。NaCl浓度为0.25 M时活性最大;在较高的渗透压值下,我们发现活性出乎意料地下降。这是首次使用pOSEX载体表达异源蛋白的报道,在仅使用NaCl或其他不渗透渗透物的生物技术过程中,它具有调节、无毒、简单和经济的优势。
{"title":"Osmoregulated TAQ polymerase gene expression in Escherichia coli.","authors":"Yeosvany Cabrera Artiles, Duniesky Martínez García, Enrique R Pérez Cruz, Gabriel J Márquez Perera, Manuel Luis Feble","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>The Thermus aquaticus DNA Polymerase I (Taq Pol I) gene was cloned into the pOSEX4 plasmid under the osmo-inducible promoter proU and subsequently expressed into the Escherichia coli MKH13 strain. The suitability of the enzyme in polymerase assays was determined in standard 35S dATP incorporation tests and by PCR. The Taq Pol I expression in this system, which is under the control of the osmotic pressure in the growth medium, was analyzed in different media and in different sodium chloride concentrations. A study of the osmolarity effects in the growth of the strain and in Taq Pol I expression shows that an increase in sodium chloride concentration limits the growth. At 0.25 M of NaCl maximum activity was observed; at higher values of osmolarity, we found an unexpected decline of activity. This is the first report of using the pOSEX vector for the expression of an heterologous protein and it is very advantageous to make a regulated, non toxic, simple and cost-effective manner of induction in a biotechnology process using just NaCl or other non-permeable osmolyte.</p>","PeriodicalId":21464,"journal":{"name":"Revista latinoamericana de microbiologia","volume":"44 1","pages":"14-8"},"PeriodicalIF":0.0,"publicationDate":"2002-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"26326842","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Graciela Castro Escarpulli, Dayrema Peña del Barrio, N Castañeda, Ailec García Azcuaga, Luis Morier Dias, Maria Guadalupe Aguilera-Arreola, Laura Bravo Farias
Fifty Aeromonas caviae strains from intestinal infection in different Cuban provinces were identified by the Aerokey II method and virulence factors were investigated. The strains did not produce haemolysins but other exoenzymes such as proteases, lipases, and DNases; additionally, all isolates adhered to the HEp-2 cell line by the Carrello method and this did not correlate with other virulence factors presence which demonstrates that the haemolysin phenotypic expression is not necessary for these strains to be pathogenic and that pathogenicity is multifactorial, each strain expressing at least one virulence factor.
{"title":"Virulence factors of A. caviae strains isolated from acute diarrheic disease in Cuba.","authors":"Graciela Castro Escarpulli, Dayrema Peña del Barrio, N Castañeda, Ailec García Azcuaga, Luis Morier Dias, Maria Guadalupe Aguilera-Arreola, Laura Bravo Farias","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>Fifty Aeromonas caviae strains from intestinal infection in different Cuban provinces were identified by the Aerokey II method and virulence factors were investigated. The strains did not produce haemolysins but other exoenzymes such as proteases, lipases, and DNases; additionally, all isolates adhered to the HEp-2 cell line by the Carrello method and this did not correlate with other virulence factors presence which demonstrates that the haemolysin phenotypic expression is not necessary for these strains to be pathogenic and that pathogenicity is multifactorial, each strain expressing at least one virulence factor.</p>","PeriodicalId":21464,"journal":{"name":"Revista latinoamericana de microbiologia","volume":"44 1","pages":"11-3"},"PeriodicalIF":0.0,"publicationDate":"2002-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"26326841","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Rolando García-Gómez, José Chávez-Espinosa, Adriana Mejía-Chávez, Carmen Duránde-Bazúa
Vegetables intake is widely recommended because of its high content of vitamins, minerals and fiber. However, the irrigation of these vegetables, using wastewaters that have received inadequate treatment often carries unseen microbial pollution that becomes a high risk potential for humans. In the present research, two of the most consumed fresh vegetables cultivated in Mexico City were analyzed, lettuce (Lactuca sativa) and Mexican coriander (Eryngium foetidum). These vegetables are commonly consumed raw. The vegetable choice and the disinfection's method were carried out by the application of two tests to two hundred people in an aleatory form. Similarly, vegetable sampling was carried out by means of a random sampling from the cultivated areas in a chosen "chinampa" (from Náhuatl or Aztec, chinamitl, bulrush or cattail stalks lattice for hydroponics cultivation). Vegetable samples were transferred, in dark plastic bags and in cool boxes at 4 +/- 1.5 degrees C, to the laboratory. Microbiological analysis for Salmonella typhi, mesophilic microorganisms, and fecal coliforms were done according to the "NOM-093SSA1-1994" (Mexico). Results obtained demonstrated that samples treated with the most preferred disinfectant, a colloidal silver based one, had a partial elimination of pathogenic microorganisms found in both vegetables lettuce (Lactuca sativa) and coriander (Eryngium foetidum) samples (mesophyllic microorganisms from 200,000 to 96,500 UFC/g and from 175,000 to 125,000 UFC/g and fecal coliforms from 75 to 0.43 NMP/g and from 150 to 2.10 NMP/g, respectively). Salmonella typhi for all samples gave a positive result. Therefore, it was recommended to the cultivators of the Xochimilco (Náhuatl or Aztec name that means "place where flowers bloom") zone, either stop using contaminated water for irrigation or to use more efficient methods in order to eliminate pathogenic microorganisms, such as diluted chlorine solutions made with commercial cotton clothing bleachers.
{"title":"Microbiological determinations of some vegetables from the Xochimilco zone in Mexico City, Mexico.","authors":"Rolando García-Gómez, José Chávez-Espinosa, Adriana Mejía-Chávez, Carmen Duránde-Bazúa","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>Vegetables intake is widely recommended because of its high content of vitamins, minerals and fiber. However, the irrigation of these vegetables, using wastewaters that have received inadequate treatment often carries unseen microbial pollution that becomes a high risk potential for humans. In the present research, two of the most consumed fresh vegetables cultivated in Mexico City were analyzed, lettuce (Lactuca sativa) and Mexican coriander (Eryngium foetidum). These vegetables are commonly consumed raw. The vegetable choice and the disinfection's method were carried out by the application of two tests to two hundred people in an aleatory form. Similarly, vegetable sampling was carried out by means of a random sampling from the cultivated areas in a chosen \"chinampa\" (from Náhuatl or Aztec, chinamitl, bulrush or cattail stalks lattice for hydroponics cultivation). Vegetable samples were transferred, in dark plastic bags and in cool boxes at 4 +/- 1.5 degrees C, to the laboratory. Microbiological analysis for Salmonella typhi, mesophilic microorganisms, and fecal coliforms were done according to the \"NOM-093SSA1-1994\" (Mexico). Results obtained demonstrated that samples treated with the most preferred disinfectant, a colloidal silver based one, had a partial elimination of pathogenic microorganisms found in both vegetables lettuce (Lactuca sativa) and coriander (Eryngium foetidum) samples (mesophyllic microorganisms from 200,000 to 96,500 UFC/g and from 175,000 to 125,000 UFC/g and fecal coliforms from 75 to 0.43 NMP/g and from 150 to 2.10 NMP/g, respectively). Salmonella typhi for all samples gave a positive result. Therefore, it was recommended to the cultivators of the Xochimilco (Náhuatl or Aztec name that means \"place where flowers bloom\") zone, either stop using contaminated water for irrigation or to use more efficient methods in order to eliminate pathogenic microorganisms, such as diluted chlorine solutions made with commercial cotton clothing bleachers.</p>","PeriodicalId":21464,"journal":{"name":"Revista latinoamericana de microbiologia","volume":"44 1","pages":"24-30"},"PeriodicalIF":0.0,"publicationDate":"2002-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"26326844","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}