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https://www.readbyqxmd.com/read/28791034/a-high-throughput-standard-pcr-based-genotyping-method-for-determining-transgene-zygosity-in-segregating-plant-populations
#1
Lige Geng, Dewayne D Deng, Martin J Wubben, Johnie N Jenkins, Jack C McCarty, Ibrokhim Abdurakhmonov
In crop research programs that implement transgene-based strategies for trait improvement it is necessary to distinguish between transgene homozygous and hemizygous individuals in segregating populations. Direct methods for determining transgene zygosity are technically challenging, expensive, and require specialized equipment. In this report, we describe a standard PCR-based protocol coupled with capillary electrophoresis that can identify transgene homozygous and hemizygous individuals in a segregating population without knowledge of transgene insertion site...
2017: Frontiers in Plant Science
https://www.readbyqxmd.com/read/28780098/prevalence-and-characterization-of-methicillin-resistant-staphylococcus-aureus-carrying-meca-or-mecc-and-methicillin-susceptible-staphylococcus-aureus-in-dairy-sheep-farms-in-central-italy
#2
G Giacinti, V Carfora, A Caprioli, D Sagrafoli, N Marri, G Giangolini, R Amoruso, M Iurescia, F Stravino, S Dottarelli, F Feltrin, A Franco, S Amatiste, A Battisti
Between January and May 2012, a total of 286 bulk tank milk samples from dairy sheep farms located in central Italy were tested for the presence of Staphylococcus aureus. One hundred fifty-three samples were positive for S. aureus (53.5%), with an average count of 2.53 log cfu/mL. A total of 679 S. aureus colonies were screened for methicillin resistance by the cefoxitin disk diffusion test, and 104 selected cefoxitin-susceptible isolates were also tested for their susceptibility to other antimicrobials representative of the most relevant classes active against Staphylococcus spp...
August 2, 2017: Journal of Dairy Science
https://www.readbyqxmd.com/read/28774859/conjugal-transfer-of-aac-6-ie-aph-2%C3%A2-ia-gene-from-native-species-and-mechanism-of-regulation-and-cross-resistance-in-enterococcus-faecalis-mcc3063-by-real-time-pcr
#3
G Jaimee, P M Halami
High level aminoglycoside resistance (HLAR) in the lactic acid bacteria (LAB) derived from food animals is detrimental. The aim of this study was to investigate the localization and conjugal transfer of aminoglycoside resistance genes, aac(6')Ie-aph(2″)Ia and aph(3')IIIa in different Enterococcus species. The cross resistance patterns in Enterococcus faecalis MCC3063 to clinically important aminoglycosides by real time PCR were also studied. Southern hybridization experiments revealed the presence of aac(6')Ie-aph(2(″))Ia and aph(3')IIIa genes conferring HLAR in high molecular weight plasmids except in Lactobacillus plantarum...
July 31, 2017: Microbial Pathogenesis
https://www.readbyqxmd.com/read/28769915/the-transient-multidrug-resistance-phenotype-of-salmonella-enterica-swarming-cells-is-abolished-by-sub-inhibitory-concentrations-of-antimicrobial-compounds
#4
Oihane Irazoki, Susana Campoy, Jordi Barbé
Swarming motility is the rapid and coordinated multicellular migration of bacteria across a moist surface. During swarming, bacterial cells exhibit increased resistance to multiple antibiotics, a phenomenon described as adaptive or transient resistance. In this study, we demonstrate that sub-inhibitory concentrations of cefotaxime, ciprofloxacin, trimethoprim, or chloramphenicol, but not that of amikacin, colistin, kanamycin or tetracycline, impair Salmonella enterica swarming. Chloramphenicol-treated S. enterica cells exhibited a clear decrease in their flagellar content, while treatment with other antibiotics that reduced swarming (cefotaxime, ciprofloxacin, and trimethoprim) inhibited polar chemoreceptor array assembly...
2017: Frontiers in Microbiology
https://www.readbyqxmd.com/read/28763019/use-of-antibiotics-for-maintenance-of-axenic-cultures-of-amphidinium-carterae-for-the-analysis-of-translation
#5
Chieh-Lun Liu, Allen R Place, Rosemary Jagus
Most dinoflagellates in culture are bacterized, complicating the quantification of protein synthesis, as well as the analysis of its regulation. In bacterized cultures of Amphidinium carterae Hulbert, up to 80% of protein synthetic activity appears to be predominantly bacterial based on responses to inhibitors of protein synthesis. To circumvent this, axenic cultures of A. carterae were obtained and shown to respond to inhibitors of protein synthesis in a manner characteristic of eukaryotes. However, these responses changed with time in culture correlating with the reappearance of bacteria...
August 1, 2017: Marine Drugs
https://www.readbyqxmd.com/read/28756809/first-report-in-china-on-the-identification-and-drug-sensitivity-of-mycobacterium-elephantis-isolated-from-the-milk-of-a-cow-with-mastitis
#6
Ling Yun Ji, Dong Lei Xu, Shu Peng Yin, Hai Can Liu, Gui Lian Li, Yi Jiang, Jian Hao Wei, Hao Zeng, Yong Liang Lou, Jian Xin Lyu, Kang Lin Wan
OBJECTIVE: In this study, milk from a cow with mastitis was analyzed to determine the presence of mycobacterial infection. Milk quality and security problems pertaining to the safe consumption of dairy products were also discussed in this study. METHODS: Milk was preprocessed with 4% NaOH. Then, mycobacteria were isolated from the milk sample on L-J medium. The isolate was identified using multiple loci Polymerase Chain Reaction (PCR) and multi-locus sequence analysis with 16S rRNA, sodA, hsp65, and ITS genes...
July 2017: Biomedical and Environmental Sciences: BES
https://www.readbyqxmd.com/read/28755354/transient-gus-gene-expression-in-date-palm-fruit-using-agroinjection-transformation-technique
#7
Mohei El-Din M Solliman, Hebaallah A Mohasseb, Abdullatif A Al-Khateeb, Jameel M Al-Khayri, Suliman A Al-Khateeb
Transient expression of foreign genes in plant tissue is a valuable tool for testing the efficacy of transformation methods. In this work, we present, for the first time, the utilization of agroinjection as an efficient transformation system for gene delivery in date palm fruit. The research utilized Agrobacterium tumefaciens strain LBA4404 harboring the binary vector pRI201-AN-GUS carrying the beta-glucuronidase (GUS) gene, under the control of a CaMV 35S and kanamycin (NPTII) as an antibiotic gene under the control of a NOS promoter...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28753625/modulation-of-kanamycin-b-and-kanamycin-a-biosynthesis-in-streptomyces-kanamyceticus-via-metabolic-engineering
#8
Wenli Gao, Zheng Wu, Junyang Sun, Xianpu Ni, Huanzhang Xia
Both kanamycin A and kanamycin B, antibiotic components produced by Streptomyces kanamyceticus, have medical value. Two different pathways for kanamycin biosynthesis have been reported by two research groups. In this study, to obtain an optimal kanamycin A-producing strain and a kanamycin B-high-yield strain, we first examined the native kanamycin biosynthetic pathway in vivo. Based on the proposed parallel biosynthetic pathway, kanN disruption should lead to kanamycin A accumulation; however, the kanN-disruption strain produced neither kanamycin A nor kanamycin B...
2017: PloS One
https://www.readbyqxmd.com/read/28750523/aptamer-embedded-zirconium-based-metal-organic-framework-composites-prepared-by-de-novo-bio-inspired-approach-with-enhanced-biosensing-for-detecting-trace-analytes
#9
Zhi-Hong Zhang, Feng-He Duan, Jia-Yue Tian, Jun-Ying He, Long-Yu Yang, Hui Zhao, Shuai Zhang, Chun-Sen Liu, Ling-Hao He, Min Chen, Di-Ming Chen, Miao Du
A series of Zr-based metal-organic framework (MOF) composites embedded with three kinds of aptamer strands (509-MOF@Apt) were achieved by a one-step de novo synthetic approach. A platform for ultrasensitive detection of analytes, namely, thrombin, kanamycin, and carcinoembryonic antigen (CEA), was also established. Considering the conformational changes caused by the binding interactions between aptamer strands and targeted molecules, the label-free electrochemical aptasensors based on 509-MOF@Apt composites could be developed to detect various target molecules...
July 28, 2017: ACS Sensors
https://www.readbyqxmd.com/read/28744538/chiral-phosphoric-acid-catalyzed-desymmetrizative-glycosylation-of-2-deoxystreptamine-and-its-application-to-aminoglycoside-synthesis
#10
Jeonghyo Lee, Alina Borovika, Yaroslav Khomutnyk, Pavel Nagorny
This work describes chiral phosphoric acid (CPA)-catalyzed desymmetrizative glycosylation of meso-diol derived from 2-deoxystreptamine. The chirality of CPA dictates the outcome of the glycosylation reactions, and the use of enantiomeric CPAs results in either C4-glycosylated (67 : 33 d.r.) or C6-glycosylated (86 : 14 d.r.) 2-deoxystreptamines. These glycosylated products can be converted to aminoglycosides, and the application of this strategy to the synthesis of protected iso-neamine and iso-kanamycin B with inverted connection at the C4 and C6 positions is described...
August 8, 2017: Chemical Communications: Chem Comm
https://www.readbyqxmd.com/read/28742090/t-dna-genome-junctions-form-early-after-infection-and-are-influenced-by-the-chromatin-state-of-the-host-genome
#11
Shay Shilo, Pooja Tripathi, Cathy Melamed-Bessudo, Oren Tzfadia, Theodore R Muth, Avraham A Levy
Agrobacterium tumefaciens mediated T-DNA integration is a common tool for plant genome manipulation. However, there is controversy regarding whether T-DNA integration is biased towards genes or randomly distributed throughout the genome. In order to address this question, we performed high-throughput mapping of T-DNA-genome junctions obtained in the absence of selection at several time points after infection. T-DNA-genome junctions were detected as early as 6 hours post-infection. T-DNA distribution was apparently uniform throughout the chromosomes, yet local biases toward AT-rich motifs and T-DNA border sequence micro-homology were detected...
July 2017: PLoS Genetics
https://www.readbyqxmd.com/read/28741151/probiotic-characteristics-of-lactobacillus-curvatus-dn317-a-strain-isolated-from-chicken-ceca
#12
Mohamed Zommiti, Nathalie Connil, Jeannette Ben Hamida, Mounir Ferchichi
The probiotic characteristics of Lactobacillus curvatus DN317, a strain isolated from chicken ceca, were evaluated. This strain was selected for study from the isolated Lactobacillus strains because it has specific anti-microbial activity against Campylobacter jejuni ATCC 33560, Camp. jejuni NCTC 11168, Listeria monocytogenes ATCC 7644, and Bacillus subtilis ATCC 8633. Lact. curvatus DN317 showed an auto-aggregation percentage of 72% and presented the highest co-aggregation with Lact. monocytogenes ATCC 7644 (68%) compared to B...
July 24, 2017: Probiotics and Antimicrobial Proteins
https://www.readbyqxmd.com/read/28734024/effect-of-solvent-and-protein-dynamics-in-ligand-recognition-and-inhibition-of-aminoglycoside-adenyltransferase-2%C3%A2-ia
#13
Valjean R Bacot-Davis, Angelia V Bassenden, Tara Sprules, Albert M Berghuis
The aminoglycoside modifying enzyme (AME) ANT(2″)-Ia is a significant target for next generation antibiotic development. Structural studies of a related aminoglycoside-modifying enzyme, ANT(3″)(9), revealed this enzyme contains dynamic, disordered, and well-defined segments that modulate thermodynamically before and after antibiotic binding. Characterizing these structural dynamics is critical for in situ screening, design, and development of contemporary antibiotics that can be implemented in a clinical setting to treat potentially lethal, antibiotic resistant, human infections...
July 21, 2017: Protein Science: a Publication of the Protein Society
https://www.readbyqxmd.com/read/28729209/clonal-spread-of-methicillin-resistant-staphylococcus-aureus-t6065-cc5-sccmecv-agrii-in-a-libyan-hospital
#14
Monia Khemiri, Ali Akrout Alhusain, Mohamed Salah Abbassi, Houyem El Ghaieb, Sofia Santos Costa, Adriana Belas, Constança Pomba, Salah Hammami
OBJECTIVES: The aim of this study was to characterize 32 MRSA isolates recovered from wound specimens of patients in a Hospital in Tripoli, Libya, during 2013. METHODS: MRSA isolates were characterized by determining their antibiotic susceptibilities, genes encoding antibiotic resistance and virulence factors, the SCCmec class, agr type, spa typing, PFGE and MLST. RESULTS: PFGE and MLST revealed that all isolates were clonal and belonged to the Clonal Complex 5 (CC5)...
July 17, 2017: Journal of Global Antimicrobial Resistance
https://www.readbyqxmd.com/read/28729208/detection-of-multidrug-resistant-mdr-bacteria-in-untreated-waste-water-disposals-of-hospitals-in-dhaka-city-bangladesh
#15
M A G Rabbani, Md Zakir Hossain Howlader, Yearul Kabir
BACKGROUND: Emergence of MDR bacteria is a global problem and a major burden for treatment of various infectious diseases. This study was performed to detect antibiotic resistant bacteria in untreated hospital waste. METHOD: Waste water samples were collected from sewerage disposal points of two renowned hospital of Dhaka city and a total of 59 Escherichia coli and 29 Klebsiella pneumonia isolates were isolated. Antibiotic susceptibility were measured by disc diffusion method...
July 17, 2017: Journal of Global Antimicrobial Resistance
https://www.readbyqxmd.com/read/28715370/inducible-t7-rna-polymerase-mediated-multigene-expression-system-pmgx
#16
Mohamed I Hassan, Fern R McSorley, Kinya Hotta, Christopher N Boddy
Co-expression of multiple proteins is increasingly essential for synthetic biology, studying protein-protein complexes, and characterizing and harnessing biosynthetic pathways. In this manuscript, the use of a highly effective system for the construction of multigene synthetic operons under the control of an inducible T7 RNA polymerase is described. This system allows many genes to be expressed simultaneously from one plasmid. Here, a set of four related vectors, pMGX-A, pMGX-hisA, pMGX-K, and pMGX-hisK, with either the ampicillin or kanamycin resistance selectable marker (A and K) and either possessing or lacking an N-terminal hexahistidine tag (his) are disclosed...
June 27, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28711791/labeled-azospirillum-brasilense-wild-type-and-excretion-ammonium-strains-in-association-with-barley-roots
#17
Adrian Richard Schenberger Santos, Rafael Mazer Etto, Rafaela Wiegand Furmam, Denis Leandro de Freitas, Karina Freire d'Eça Nogueira Santos, Emanuel Maltempi de Souza, Fábio de Oliveira Pedrosa, Ricardo Antônio Ayub, Maria Berenice Reynaud Steffens, Carolina Weigert Galvão
Soil bacteria colonization in plants is a complex process, which involves interaction between many bacterial characters and plant responses. In this work, we labeled Azospirillum brasilense FP2 (wild type) and HM053 (excretion-ammonium) strains by insertion of the reporter gene gusA-kanamycin into the dinitrogenase reductase coding gene, nifH, and evaluated bacteria colonization in barley (Hordeum vulgare). In addition, we determined inoculation effect based on growth promotion parameters. We report an uncommon endophytic behavior of A...
July 4, 2017: Plant Physiology and Biochemistry: PPB
https://www.readbyqxmd.com/read/28710637/stabilization-of-enzymes-by-using-thermophiles
#18
Ana Luisa Ribeiro, Mercedes Sánchez, Aurelio Hidalgo, José Berenguer
Manufactured steroid compounds have many applications in the pharmaceutical industry. Due to the chemical complexity and chirality of steroids, there is an increasing demand for enzyme-based bioconversion processes to replace those based on chemical synthesis. In this context, thermostability of the involved enzymes is a highly desirable property as both the increased half-life of the enzyme and the enhanced solubility of substrates and products will improve the yield of the reactions. Metagenomic libraries from thermal environments are potential sources of thermostable enzymes of prokaryotic origin, but the number of expected hits could be quite low for enzymes handling substrates such as steroids, rarely found in prokaryotes...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28705770/in%C3%A2-vitro-activity-of-aminoglycosides-clofazimine-d-cycloserine-and-dapsone-against-83-mycobacterium-avium-complex-clinical-isolates
#19
Chen-Cheng Huang, Ming-Feng Wu, Hui-Chen Chen, Wei-Chang Huang
BACKGROUND/PURPOSE: Treatment success rates for Mycobacterium avium complex (MAC) diseases range from 50% to 55% only. To explore effective antimicrobials against either Mycobacteriumintracellulare or M. avium, we determined in vitro activities of five aminoglycosides, clofazimine, dapsone and d-cycloserine compared with primary (clarithromycin) and secondary (moxifloxacin and linezolid) antimycobacterial agents. METHODS: 83 non-duplicate clinical MAC isolates were collected from sputum and identified at the species level by PCR and restriction enzyme analysis of the 65 kDa hsp and rpoB genes...
June 28, 2017: Journal of Microbiology, Immunology, and Infection, Wei Mian Yu Gan Ran za Zhi
https://www.readbyqxmd.com/read/28698044/resistance-to-first-and-second-line-antituberculosis-drugs-in-southern-taiwan-implications-for-empirical-treatment
#20
Cheng-Yu Kuo, Wen-Hung Wang, Chung-Hao Huang, Yen-Hsu Chen, Po-Liang Lu
BACKGROUND: Multidrug-resistant and extensively drug-resistant tuberculosis infections cause public health concerns worldwide. Local epidemiologic data about the drug resistance of Mycobacteriumtuberculosis isolate (Mtb) is critical to guide appropriate empirical therapy to cure patients and restrain the spread of tuberculosis. METHODS: Antituberculosis susceptibility testing was performed for 287 Mtbs, including 63 MDR-Mtbs collected in southern Taiwan from 2011 to 2015...
June 29, 2017: Journal of Microbiology, Immunology, and Infection, Wei Mian Yu Gan Ran za Zhi
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