Characterization of Nosocomial Strains of Enterobacter aerogenes by Arbitrarily Primed-PCR Analysis and Ribotyping

1995 ◽  
Vol 16 (4) ◽  
pp. 224-230 ◽  
Author(s):  
Florence Grattard ◽  
Bruno Pozzetto ◽  
Lionel Tabard ◽  
Marianne Petit ◽  
Alain Ros ◽  
...  
Keyword(s):  
Enterobacter Aerogenes ◽  
Pcr Analysis ◽  
Arbitrarily Primed Pcr

Characterization of Nosocomial Strains of Enterobacter aerogenes by Arbitrarily Primed-PCR Analysis and Ribotyping

1995 ◽  
Vol 16 (4) ◽  
pp. 224-230 ◽  
Author(s):  
Florence Grattard ◽  
Bruno Pozzetto ◽  
Lionel Tabard ◽  
Marianne Petit ◽  
Alain Ros ◽  
...  
Keyword(s):  
Enterobacter Aerogenes ◽  
Pcr Analysis ◽  
Arbitrarily Primed Pcr

scholarly journals Identification of Entomopathogenic Nematode-Associated Bacteria Originating from Mojokerto

2021 ◽  
Vol 5 (1) ◽  
pp. 31-38
Author(s):  
Theresia Desy Askitosari ◽  
Tjandra Pantjajani ◽  
Stephanie Nathania ◽  
Amelia Fedoragnes Wahyudi ◽  
Nancy Christina Sugianto
Keyword(s):  
Pest Control ◽  
Entomopathogenic Nematode ◽  
Enterobacter Aerogenes ◽  
Symbiotic Bacteria ◽  
Pcr Analysis ◽  
Rrna Gene ◽  
Biochemical Tests ◽  
Gram Negative ◽  
Natural Pest Control

Entomopathogenic nematodes (EPN) are one of the soil worms that have been widely used as a natural pest control. EPN has its pathogenic capability because of the mutualistic interactions between nematodes and symbiotic bacteria inside the digestive tract of nematodes. Symbiotic bacteria capable of producing exoenzymes that are toxic to insects. The isolation of symbiotic bacteria accomplished by infection of obtained EPN (Belik II isolate) into Tenebrio molitor larvae. Symbiotic bacteria were isolated from the hemolymph of dead larvae on NBTA media. Isolation of symbiotic bacteria was successfully obtained two morphologically distinct bacteria: B 3.1 isolate and B 4 isolate. Both bacteria were further identified using PCR analysis of the 16S rRNA gene. Based on the sequencing results, the B 3.1 isolate was in accordance with Acinetobacter pittii strain ATCC 19004, while B 4 isolate was in accordance with bacteria Enterobacter aerogenes strain KCTC 2190. The characterization of B 3.1 isolate was shown to have similarities with Acinetobacter sp., i.e.: gram-negative, non - motile, rod-shaped, and some other characteristics of biochemical tests. While the characterization of B 4 isolate was shown to have similarities with E. aerogenes i.e.: gram-negative, rod-shaped, motile, and some other characteristics of biochemical tests. These findings will be the potential to be applied as biological agents in pest control.

Characterization of Partial Gene Deletions in Type III von Willebrand Disease with Alloantibody Inhibitors

1994 ◽  
Vol 72 (02) ◽  
pp. 180-185 ◽  
Author(s):  
David J Mancuso ◽  
Elodee A Tuley ◽  
Ricardo Castillo ◽  
Norma de Bosch ◽  
Pler M Mannucci ◽  
...  
Keyword(s):  
Von Willebrand Factor ◽  
Von Willebrand Disease ◽  
Pcr Analysis ◽  
Gene Deletions ◽  
Factor Gene ◽  
Type Iii ◽  
Large Deletions ◽  
Von Willebrand ◽  
Willebrand Factor

Summaryvon Willebrand factor gene deletions were characterized in four patients with severe type III von Willebrand disease and alloantibodies to von Willebrand factor. A PCR-based strategy was used to characterize the boundaries of the deletions. Identical 30 kb von Willebrand factor gene deletions which include exons 33 through 38 were identified in two siblings of one family by this method. A small 5 base pair insertion (CCTGG) was sequenced at the deletion breakpoint. PCR analysis was used to detect the deletion in three generations of the family, including two family members who are heterozygous for the deletion. In a second family, two type III vWD patients, who are distant cousins, share an -56 kb deletion of exons 22 through 43. The identification and characterization of large vWF gene deletions in these type III vWD patients provides further support for the association between large deletions in both von Willebrand factor alleles and the development of inhibitory alloantibodies.

Acanthamoeba Species in Tap Water, Egypt

2017 ◽  
Vol 9 (1) ◽  
Author(s):  
Ahmad Z Al-Herrawy ◽  
Mohamed A Marouf ◽  
Mahmoud A. Gad
Keyword(s):  
Water Samples ◽  
Tap Water ◽  
Pcr Analysis ◽  
Pulmonary Infections ◽  
Clinical Syndromes ◽  
Acanthamoeba Species ◽  
Amebic Encephalitis ◽  
Acanthamoeba Culbertsoni ◽  
Autumn And Winter

Genus Acanthamoeba causes 3 clinical syndromes amebic keratitis, granulomatous amebic encephalitis and disseminated granulomatous amebic disease (eg, sinus, skin and pulmonary infections). A total of 144 tap water samples were collected from Giza governorate, Egypt. Samples were processed for detection of Acanthamoeba species using non-nutrient agar (NNA) and were incubated at 30oC. The isolates of Acanthamoeba were identified to species level based on the morphologic criteria. Molecular characterization of the Acanthamoeba isolates to genus level was performed by using PCR. The obtained results showed that the highest occurrence percentage of Acanthamoeba species in water samples was observed in summer season (38.9%), then it decreased to be 30.6% in spring and 25% in each of autumn and winter. PCR analysis showed that 100% of 43 Acanthamoeba morphologically positive samples were positive by genus specific primer. In the present study eight species of Acanthamoeba can be morphologically recognized namely Acanthamoeba triangularis, Acanthamoeba echinulata, Acanthamoeba astronyxis, Acanthamoeba comandoni, Acanthamoeba griffini, Acanthamoeba culbertsoni, Acanthamoeba quina and Acanthamoeba lenticulata. In conclusion, the most common Acanthamoeba species in tap water was Acanthamoeba comandoni

scholarly journals Genome-Wide Identification and Characterization of Hsf and Hsp Gene Families and Gene Expression Analysis under Heat Stress in Eggplant (Solanum melongema L.)

Horticulturae ◽  
2021 ◽  
Vol 7 (6) ◽  
pp. 149
Author(s):  
Chao Gong ◽  
Qiangqiang Pang ◽  
Zhiliang Li ◽  
Zhenxing Li ◽  
Riyuan Chen ◽  
...  
Keyword(s):  
Heat Stress ◽  
Gene Families ◽  
High Temperature Stress ◽  
Pcr Analysis ◽  
Rna Seq ◽  
Genome Wide ◽  
Motif Composition ◽  
Hsp Genes ◽  
Identification And Characterization

Under high temperature stress, a large number of proteins in plant cells will be denatured and inactivated. Meanwhile Hsfs and Hsps will be quickly induced to remove denatured proteins, so as to avoid programmed cell death, thus enhancing the thermotolerance of plants. Here, a comprehensive identification and analysis of the Hsf and Hsp gene families in eggplant under heat stress was performed. A total of 24 Hsf-like genes and 117 Hsp-like genes were identified from the eggplant genome using the interolog from Arabidopsis. The gene structure and motif composition of Hsf and Hsp genes were relatively conserved in each subfamily in eggplant. RNA-seq data and qRT-PCR analysis showed that the expressions of most eggplant Hsf and Hsp genes were increased upon exposure to heat stress, especially in thermotolerant line. The comprehensive analysis indicated that different sets of SmHsps genes were involved downstream of particular SmHsfs genes. These results provided a basis for revealing the roles of SmHsps and SmHsp for thermotolerance in eggplant, which may potentially be useful for understanding the thermotolerance mechanism involving SmHsps and SmHsp in eggplant.

scholarly journals FlrA, a σ54-Dependent Transcriptional Activator in Vibrio fischeri, Is Required for Motility and Symbiotic Light-Organ Colonization

2003 ◽  
Vol 185 (12) ◽  
pp. 3547-3557 ◽  
Author(s):  
Deborah S. Millikan ◽  
Edward G. Ruby
Keyword(s):  
Vibrio Fischeri ◽  
Transcriptional Activator ◽  
Pcr Analysis ◽  
Euprymna Scolopes ◽  
Wild Type ◽  
Flagellar Regulon ◽  
Colonization Factors ◽  
Recognition Motifs ◽  
Arbitrarily Primed Pcr ◽  
Organ Colonization

ABSTRACT Flagellum-mediated motility of Vibrio fischeri is an essential factor in the bacterium's ability to colonize its host, the Hawaiian squid Euprymna scolopes. To begin characterizing the nature of the flagellar regulon, we have cloned a gene, designated flrA, from V. fischeri that encodes a putative σ54-dependent transcriptional activator. Genetic arrangement of the flrA locus in V. fischeri is similar to motility master-regulator operons of Vibrio cholerae and Vibrio parahaemolyticus. In addition, examination of regulatory regions of a number of flagellar operons in V. fischeri revealed apparent σ54 recognition motifs, suggesting that the flagellar regulatory hierarchy is controlled by a similar mechanism to that described in V. cholerae. However, in contrast to its closest known relatives, flrA mutant strains of V. fischeri ES114 were completely abolished in swimming capability. Although flrA provided in trans restored motility to the flrA mutant, the complemented strain was unable to reach wild-type levels of symbiotic colonization in juvenile squid, suggesting a possible role for the proper expression of FlrA in regulating symbiotic colonization factors in addition to those required for motility. Comparative RNA arbitrarily primed PCR analysis of the flrA mutant and its wild-type parent revealed several differentially expressed transcripts. These results define a regulon that includes both flagellar structural genes and other genes apparently not involved in flagellum elaboration or function. Thus, the transcriptional activator FlrA plays an essential role in regulating motility, and apparently in modulating other symbiotic functions, in V. fischeri.

scholarly journals Extended-Spectrum β-Lactamase-Producing Enterobacteriaceae in Community and Private Health Care Centers

2003 ◽  
Vol 47 (11) ◽  
pp. 3506-3514 ◽  
Author(s):  
Corinne Arpin ◽  
Véronique Dubois ◽  
Laure Coulange ◽  
Catherine André ◽  
Isabelle Fischer ◽  
...  
Keyword(s):  
Nursing Home ◽  
Dna Typing ◽  
Enterobacter Aerogenes ◽  
Pcr Analysis ◽  
Extended Spectrum ◽  
The Family ◽  
Ambulatory Patients ◽  
Providencia Stuartii ◽  
Plasmid Dissemination ◽  
Esbl Producers

ABSTRACT In 1999, 39 of 2,599 isolates of the family Enterobacteriaceae (1.5%) collected by eight private laboratories in the Aquitaine region in France produced an extended-spectrum β-lactamase (ESBL). Among these were 19 Enterobacter aerogenes isolates; 8 Klebsiella pneumoniae isolates; 6 Escherichia coli isolates; 3 Proteus mirabilis isolates; and 1 isolate each of Serratia marcescens, Morganella morganii, and Providencia stuartii. ESBL producers were isolated from 38 patients, including 33 residents of 11 clinics or nursing homes and 5 ambulatory patients. Seven different ESBLs were characterized. These mainly consisted of TEM-24 (25 isolates) and TEM-21 (9 isolates), but TEM-15 (2 isolates) and TEM-3, TEM-19, SHV-4, and CTX-M-1 (1 isolate each) were also characterized. Seven strains showed the coexistence of different TEM- and/or SHV-encoding genes, including a new SHV-1 variant, SHV-44, defined by the substitution R205L previously reported for SHV-3 in association with S238G. The epidemiology of the ESBL producers was investigated by random amplification of polymorphic DNA, typing by enterobacterial repetitive intergenic consensus PCR, analysis of resistance cotransferred with the ESBL, and analysis of the restriction profiles of the ESBL-encoding plasmids. Of the TEM-24-expressing strains, 18 were E. aerogenes isolates, including 9 from the same clinic, that were representatives of the epidemic clone disseminating in France. Of the TEM-21-producing strains that belonged to different species of the family Enterobacteriaceae (E. coli, K. pneumoniae, and P. mirabilis), 8 were isolated in the same nursing home. Outbreaks due to strain and/or plasmid dissemination in these clinic and nursing home were demonstrated. The presence of ESBL producers in five ambulatory patients probably resulted from nosocomial acquisition. Our data highlight the serious need to monitor patients for ESBL-producing Enterobacteriaceae in general practice.

Characterization of alpha-adrenoceptor gene expression in arterial and venous smooth muscle

1993 ◽  
Vol 265 (5) ◽  
pp. H1501-H1509 ◽  
Author(s):  
P. Ping ◽  
J. E. Faber
Keyword(s):  
Smooth Muscle ◽  
Messenger Rna ◽  
Vena Cava ◽  
Pcr Analysis ◽  
Relative Distribution ◽  
Alpha Adrenoceptor ◽  
Aortic Media ◽  
Unique Restriction ◽  
Polymerase Chain

Six genes coding for three unique alpha 1- (1A, 1B, 1C) and three unique alpha 2- (2A, 2B, 2C) adrenergic receptor (AR) subtypes have been cloned. Ligand binding and contractile studies have demonstrated that both alpha 1- and alpha 2-ARs can exist on vascular smooth muscle (VSM) cells, although less is known about the relative distribution and specific subtypes in different vascular segments. In the present study polymerase chain reaction (PCR) analysis was used to characterize the species of alpha-AR messenger RNA (mRNA) present in freshly isolated rat thoracic aortic media and vena cava and in cultured VSM cells (passage 2) derived from both sources. To prevent possible contamination of VSM mRNA, aortic media was separated from adventitia, and vessels were denuded of endothelial cells. Oligonucleotide primers specific for each of the six adrenergic genes were synthesized and used to probe for the presence of alpha-AR mRNA species after reverse transcription of total cellular RNA to cDNA. PCR-amplified AR transcripts were distinguished by the size of amplified DNA fragments and unique restriction endonuclease cleavage. Expression of alpha 1C- or alpha 2C-mRNA was not detected in vascular tissues or cultured VSM cells, although the alpha 2C-primers detected the expected alpha 2C expression in cerebral cortex. Only alpha 1A-mRNA was detected in aortic adventitia. VSM from aorta expressed alpha 1A-, alpha 1B-, and alpha 2A-mRNA, and this pattern was preserved in cultured aortic VSM. Vena cava also expressed both alpha 1A and alpha 1B; however only alpha 2B-mRNA was detected.(ABSTRACT TRUNCATED AT 250 WORDS)

scholarly journals Identification and Functional Characterization of the CVOMT and EOMT Genes Promoters from Ocimum Basilicum L

2021 ◽  
Author(s):  
Fatemeh Khakdan ◽  
Zahra Shirazi ◽  
Mojtaba Ranjbar
Keyword(s):  
Water Deficit ◽  
Transcriptional Control ◽  
Functional Characterization ◽  
Pcr Analysis ◽  
Water Deficit Stress ◽  
Specific Expression ◽  
Stress Dependent ◽  
First Time ◽  
Dependent Mechanism

Abstract Methyl chavicol and methyl eugenol are important phenylpropanoid compounds previously purified from basil. These compounds are significantly enhanced by the water deficit stress-dependent mechanism. Here, for the first time, pObCVOMT and pObEOMT promoters were extracted by the genome walking method. They were then cloned into the upstream of the β-glucuronidase (GUS) reporter gene to identify the pattern of GUS water deficit stress-specific expression. Histochemical GUS assays showed in transgenic tobacco lines bearing the GUS gene driven by pObCVOMT and pObEOMT promoters, GUS was strongly expressed under water deficit stress. qRT-PCR analysis of pObCVOMT and pObEOMT transgenic plants confirmed the histochemical assays, indicating that the GUS expression is also significantly induced and up-regulated by increasing density of water deficit stress. This indicates these promoters are able to drive inducible expression. The cis-acting elements analysis showed that the pObCVOMT and pObEOMT promoters contained dehydration or water deficit-related transcriptional control elements.

scholarly journals Use of "EP"(Peroxidase) allele in soybean varietal characterization

2014 ◽  
Vol 36 (4) ◽  
pp. 465-470
Author(s):  
Bárbara Panoff Valário ◽  
Cláudio Cavariani ◽  
José de Barros França-Neto ◽  
Elisa Serra Negra Vieira ◽  
Juliana Pereira Bravo ◽  
...  
Keyword(s):  
Plant Production ◽  
Pcr Analysis ◽  
Agarose Gel Electrophoresis ◽  
Conventional Pcr ◽  
Chain Reaction ◽  
Pcr Assay ◽  
Molecular Biology Technique ◽  
Soybean Cultivars ◽  
Colorimetric Test

The aim of this work was to evaluate the use of the molecular biology technique of PCR (Polimerase Chain Reaction) in the characterization of soybean cultivars. The study was performed at the Department of Plant Production, Faculty of Agricultural Sciences/ UNESP and Institute of Bioscience, Botucatu-SP. Fourteen commercial soybean cultivars were used, of which six were selected as positive reaction to peroxidase (BRS 320, BRS 284, BRS 232, BRS 7860RR, BRSMG 760SRR, BRS295RR), four as negative reaction (BRS 326, BRS 8160RR, BRSMG 800A (NutriSoy), BRS Valiosa RR) and four as double reaction (BRSGO 8060, BRS 270RR, FTS Campo Mourão and BRS 239). Thus, the results attained by the traditional biochemical colorimetric test for the 14 cultivars were compared with the conventional PCR assay. For PCR analysis, DNA was extracted from whole seeds and the primers were tested, and subsequently PCR and agarose gel electrophoresis were performed. The combination of primers prx9 + prx10 confirmed the use of the PCR reaction to characterize soybean cultivars considered doubtful by conventional colorimetric text.

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