Consulta: subjectFacets:"Plasmid"
Registros recuperados: 13
Data/hora: 04/06/2024 01:37:55

An improved system for competent cell preparation and high efficiency plasmid transformation using different Escherichia coli strains
Provedor de dados: 69
Autores: Tu,Zhiming; He,Guangyuan; Li,Kexiu X.; Chen,Mingjie J.; Chang,Junli; Chen,Ling; Yao,Qing; Liu,Dongping P.; Ye,Huan; Shi,Jiantao; Wu,Xuqian .
This paper describes an efficient bacterial transformation system that was established for the preparation of competent cells, plasmid preparation, and for the storage in bacterial stocks in our laboratory. Using this method, a number of different plasmids have been amplified for further experiments. Competent cells for bacterial transformation were prepared by the calcium chloride method with an optimum concentration of 75 mM. Three different strains of Escherichia coli that were tested are DH5&amp;#945;, TG1 and XL1 blue, and the most efficient strain being XL1 blue. The optimal optical density (OD600) range for competent cell preparation varied for each of the strains investigated, and for XL1 blue it was 0.15-0.45; for TG1 it was 0.2-0.5; and for...
Tipo: Journal article
Palavras-chave: Competent cells; E. coli; Plasmid; Storage; Transformation.
Ano: 2005
URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582005000100014

Cre-loxP recombination vectors for promoter studies
Provedor de dados: 69
Autores: Pedersen,Nina; Tuxen Poulsen,Thomas; Skovgaard Poulsen,Hans.
For promoter studies the cloning, subcloning and transfer to different plasmid vectors usually requires use of restriction enzymes and ligation reactions. One obstacle is the nucleotide polymorphisms of eukaryotic genomic DNA, which has the consequence that a sequence often differs from published sequences. Therefore sequencing, rigorous restriction enzyme analysis or introduction of suitable sites has to be performed prior to cloning and subcloning. In addition, conventional methods using restriction enzymes, insert purifications and ligations is expensive and labour demanding. We have developed a fast, efficient and inexpensive Cre recombinase-loxP based method, which allows cloning of promoter regions and subcloning of these into a variety of vectors in...
Tipo: Journal article
Palavras-chave: Cloning; Cre recombinase; Plasmid; Reporter genes; Therapeutic genes.
Ano: 2007
URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582007000200013

Degradation of methomyl by the novel bacterial strain Stenotrophomonas maltophilia M1
Provedor de dados: 69
Autores: Mohamed,Mervat S.
The use of microorganisms in the degradation and detoxification of many toxic xenobiotics, especially pesticides, is an efficient tool for the decontamination of polluted sites in the environment. A novel bacterial strain (M1) was isolated from several water samples contaminated with methomyl which is capable of degrading methomyl pesticide (1000 ppm) in the presence of 0.05% glucose. These water samples were collected from different irrigation sites in Egypt where methomyl is heavily applied. The partial sequence of 16SrRNA gene of the isolate showed the highest similarity to Stenotrophomonas maltophilia. Restriction fragment patterns of isolated plasmid DNA showed that this strain harbours two different plasmids PMa (8Kb) and PMb (5Kb). PMb succeeded to...
Tipo: Journal article
Palavras-chave: Carbamate; Methomyl; Plasmid; SPE-LC-ESI-MS; Stenotrophomonas.
Ano: 2009
URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582009000400006

Deletion of DNA sequences of using a polymerase chain reaction based approach
Provedor de dados: 69
Autores: Pérez-Pinera,Pablo; Menéndez-González,Manuel; Vega,José Antonio.
We developed a simple, rapid and reliable method to delete DNA fragments in plasmids using a polymerase chain reaction based amplification of the circular DNA sequence that excludes the fragment to be deleted. The primers are designed to contain a non-complementary 5' sequence consisting of a restriction enzyme target sequence. Following PCR amplification, the plasmid is digested with Dpn I to eliminate the template DNA, with the chosen restriction enzyme, and ligated. The only limitation is the selection of the restriction enzyme target sequence that must not be present in the original plasmid. The method is straightforward in its execution and success relies on a meticulous primer design that permits us obtain 100% of transformants containing the desired...
Tipo: Journal article
Palavras-chave: Deletion; Mutagenesis; Polymerase chain reaction; Plasmid.
Ano: 2006
URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582006000500018

Frequency of transferable multiple antibiotic resistance amongst coliform bacteria isolated from a treated sewage effluent in Antofagasta, Chile
Provedor de dados: 69
Autores: Silva,Juan; Castillo,Gabriela; Callejas,Lorena; López,Héctor; Olmos,Janet.
Antibiotic-resistant coliform bacteria from raw and treated water from activated sludge of a sewage treatment plant were isolated on eosin methylene blue agar (EMB) plates in the separate presence of ampicillin, tetracycline, chloramphenicol and streptomycin. Antibiotic resistant coliforms obtained from inflow and effluent waters were studied to determine influences of treatment on the species composition, resistance patterns and transferable resistance in these bacteria. The bacterial counts obtained showed a high proportion of antibiotic resistant coliforms in the treated effluent. The resistant coliform species were eliminated at different rates by the treatment. Escherichia coli decreased, whereas the proportion of Klebsiella sp increased. The...
Tipo: Journal article
Palavras-chave: Antibiotic resistance; Coliforms; Plasmid; Sewage treatment; Water pollution.
Ano: 2006
URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582006000500008

Interaction de la bactérie Vibrio harveyi avec son hôte, l'hormeau Haliotis tuberculata : approches physiologiques, cellulaires et moléculaires
Provedor de dados: 5
Autores: Travers, Marie-agnes.
Abalone aquaculture only recently developed in western Europe, but already it has to face numerous pathologies. Since 1998, the natural and farm stocks of Haliotis tuberculata suffer regularly high mortalities, mainly due to the pathogenic bacterium, Vibrio harveyi. The vibriosis of the European abalone, in particular its macroscopic signs, and the conditions necessary for its development have been studied in detail by field and laboratory experiments. We have been able to show the importance of the interplay between water temperature (&gt;17°C), animal physiology (reproduction period) and a pathogenic strain (harbouring the plasmid pVCR1) for the development of this abalone disease. A limited localisation study of the abalone target tissues (gills, muscle...
Tipo: Text
Palavras-chave: Ormeau; Haliotis tuberculata; Bactéries; Vibrio harveyi; Vibriose; Température; Immunité; Reproduction; Hémocytes; MAP Kinases; Plasmide; Virulence; GFP; Abalone; Haliotis tuberculata; Bacteria; Vibrio harveyi; Vibriosis; Temperature; Immunity; Reproduction; Haemocytes; MAP Kinases; Plasmid; Virulence; GFP.
Ano: 2008
URL: http://archimer.ifremer.fr/doc/00176/28729/27196.pdf

Development of TaqMan real-time PCR assays for monitoring Vibrio harveyi infection and a plasmid harbored by virulent strains in European abalone Haliotis tuberculata aquaculture
Provedor de dados: 5
Autores: Schikorski, David; Renault, Tristan; Paillard, Christine; Bidault-toffin, A.; Tourbiez, Delphine; Saulnier, Denis.
The Gram-negative bacterium Vibrio harveyi is known to be highly pathogenic for the European abalone Haliotis tuberculata, which is a gastronomically important marine gastropod with a high commercial value. Since 1998, some particular bacterial strains are described as implicated in recurrent mortality outbreaks in French farm and field stocks of abalone. Recently, a 9.6kb plasmid named pVCR1, was shown to be harbored by one highly V. harveyi virulent ORM4 strain suggesting its involvement in virulence phenotype. Thus, we have developed in the present study two TaqMan real-time PCR assays allowing to (i) rapidly and specifically detect, by a duplex procedure and in less than 2 h, both V. harveyi and the presence of plasmid pVCR1 from unidentified bacterial...
Tipo: Text
Palavras-chave: Vibrio harveyi; Haliotis tuberculata; Real-time PCR; Plasmid; Marine pathogen; Molecular diagnostic.
Ano: 2013
URL: http://archimer.ifremer.fr/doc/00124/23540/21383.pdf

Plasmids in Mycoplasma species isolated from goats and sheep and their preliminary typing
Provedor de dados: 59
Autores: Nascimento,Elmiro R.; DaMassa,Al J.; Yamamoto,Richard; Nascimento,M. Graça F.
One-hundred-five (105) clinical isolates of mycoplasma from caprine origin and one isolate from ovine were surveyed for plasmids, which were present in thirty-three (31%) of them. These mycoplasmas originated from 13 herds. Ten of them were symptomatic for mycoplasmal disease (mastitis, polyarthritis, septicemia) and three herds were asymptomatic, i.e., clinically normal. Twenty-eight isolates were Mycoplasma mycoides subspecies mycoides LC (large colony or caprine biotype), four were Mycoplasma capricolum subsp. capricolum and one was Mycoplasma cottewii. The isolated plasmids were linearized by EcoRI, EcoRV, EcoRI and EcoRV or BamHI and EcoRV, and were of five sizes (1.1, 1.6, 1.7, 1.8, and 1.9 Kbp). Based on restriction enzyme digestion and size of the...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: Mycoplasma; Plasmid; Goat; Endonuclease; Electrophoresis.
Ano: 1999
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0001-37141999000100006

Effects of fenoprofen on the labeling of blood constituents with technetium-99m, the morphology of red blood cells and the plasmid
Provedor de dados: 52
Autores: Pereira,Marcia de Oliveira; Rocha,Gabrielle de Souza; Lombardi,Simone dos Santos; Geller,Mauro; Pereira,Mário José; Santos-Filho,Sebastião David; Fonseca,Adenilson de Souza da; Bernardo-Filho,Mario.
The aim of this work was to evaluate the effect of fenoprofen on the labeling of blood constituents with technetium-99m, on the morphology of red blood cells and on the plasmid DNA. Blood samples from Wistar rats were incubated with fenoprofen and the assay of labeling of blood constituents with technetium-99m (99mTc) was performed. Blood cells, plasma, soluble and insoluble fractions of blood cells and plasma were separated. The radioactivity in each fraction was counted and percentage of incorporated radioactivity (%ATI) was determined. Blood smears were prepared, fixed, stained and the qualitative and quantitative morphology of the red blood cells (RBC) was evaluated. Plasmid (pBSK) was incubated with fenoprofen with stannous chloride, and agarose gel...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: Technetium-99m; Blood; Morphology; Plasmid; Fenoprofen.
Ano: 2008
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132008000700022

Purification of plasmid (pVaxLacZ) by hydrophobic interaction chromatography
Provedor de dados: 52
Autores: Moreira,Keila Aparecida; Diogo,Margarida; Prazeres,Duarte Miguel; Lima Filho,José Luiz de; Porto,Ana Lúcia Figueiredo.
This paper describes a method for the plasmid DNA purification, which includes an ammonium sulphate precipitation, followed by hydrophobic interaction chromatography (HIC) using Phenyl Sepharose 6 Fast Flow (low sub). The use of HIC took advantage of the more hydrophobic character of single stranded nucleic acid impurities as compared with double-stranded plasmid DNA.
Tipo: Info:eu-repo/semantics/article
Palavras-chave: Gene therapy; Hydrophobic interaction chromatography; Plasmid; Purification.
Ano: 2005
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132005000400014

DNA vaccines for viral diseases
Provedor de dados: 56
Autores: Donnelly,J.J.; Ulmer,J.B.
DNA plasmids encoding foreign proteins may be used as immunogens by direct intramuscular injection alone, or with various adjuvants and excipients, or by delivery of DNA-coated gold particles to the epidermis through biolistic immunization. Antibody, helper T lymphocyte, and cytotoxic T lymphocyte (CTL) responses have been induced in laboratory and domesticated animals by these methods. In a number of animal models, immune responses induced by DNA vaccination have been shown to be protective against challenge with various infectious agents. Immunization by injection of plasmids encoding foreign proteins has been used successfully as a research tool. This review summarizes the types of DNA vaccine vectors in common use, the immune responses and protective...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: Plasmid; Nucleic acid; Protection; Antibodies; T cells; CTL.
Ano: 1999
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X1999000200010

Comparative study of wild and transformed salt tolerant bacterial strains on Triticum aestivum growth under salt stress
Provedor de dados: 58
Autores: Afrasayab,Shazia; Faisal,Muhammad; Hasnain,Shahida.
Eleven salt tolerant bacteria isolated from different sources (soil, plants) and their transformed strains were used to study their influence on Triticum aestivum var. Inqlab-91 growth under salt (100 mM NaCl) stress. Salt stress caused reduction in germination (19.4%), seedling growth (46%) and fresh weight (39%) in non-inoculated plants. In general, both wild and transformed strains stimulated germination, seedling growth and fresh weight in salt free and salt stressed conditions. At 100 mM NaCl, Staphylococcus xylosus ST-1 caused 25% increments in seedling length over respective control. Soluble protein content significantly enhanced (49%) under salt stress as compared to salt free control. At 100 mM NaCl parental strain PT-5 resulted about 32%...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: Triticum aestivum; Plasmid; Rhizobacteria; Salinity; Auxin; Transformation.
Ano: 2010
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822010000400013

pMEX01, a 70 kb plasmid isolated from Escherichia coli that confers resistance to multiple β-lactam antibiotics
Provedor de dados: 58
Autores: Hernandez-Flores,Jose Luis; Pérez,Juan Caballero; Gutiérrez,Carlos Saldaña; Hernández,Andrés Cruz; Alonso,Gerardo Soto; Hernández,Sergio Pacheco; Gómez,Sergio Romero; Fernández,Francisco; Loske,Achim M.; Guillén,Juan Campos.
Abstract Multidrug-resistant microorganisms are of great concern to public health. Genetic mobile elements, such as plasmids, are among the most relevant mechanisms by which bacteria achieve this resistance. We obtained an Escherichia coli strain CM6, isolated from cattle presenting severe diarrheic symptoms in the State of Querétaro, Mexico. It was found to contain a 70 kb plasmid (pMEX01) with a high similarity to the pHK01-like plasmids that were previously identified and described in Hong Kong. Analysis of the pMEX01 sequence revealed the presence of a blaCTX-M-14 gene, which is responsible for conferring resistance to multiple β-lactam antibiotics. Several genes putatively involved in the conjugative transfer were also identified on the plasmid. The...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: Escherichia coli; Plasmid; Antibiotic resistance; BlaCTX-M-14 gene.
Ano: 2018
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822018000300569