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https://www.readbyqxmd.com/read/28074221/exploring-the-thermostable-properties-of-halohydrin-dehalogenase-from-agrobacterium-radiobacter-ad1-by-a-combinatorial-directed-evolution-strategy
#1
Zhiyun Wu, Wenfeng Deng, Yapei Tong, Qian Liao, Dongmin Xin, Huashun Yu, Juan Feng, Lixia Tang
As a crucial factor for biocatalysts, protein thermostability often arises from a combination of factors that are often difficult to rationalize. In this work, the thermostable nature of halohydrin dehalogenase from Agrobacterium radiobacter AD1 (HheC) was systematically explored using a combinatorial directed evolution approach. For this, a mutagenesis library of HheC mutants was first constructed using error-prone PCR with low mutagenesis frequency. After screening approximately 2000 colonies, six mutants with eight mutation sites were obtained...
January 10, 2017: Applied Microbiology and Biotechnology
https://www.readbyqxmd.com/read/28069822/transcriptional-activation-of-virulence-genes-of-rhizobium-etli
#2
Luyao Wang, Benoît Lacroix, Jianhua Guo, Vitaly Citovsky
Recently, Rhizobium etli has emerged, in addition to Agrobacterium spp., as a prokaryotic species that encodes a functional machinery for DNA transfer to plant cells. To understand this R. etli-mediated genetic transformation, it would be useful to define how its vir genes respond to the host plants. Here, we explored the transcriptional activation of the vir genes contained on the R. etli p42a plasmid. Using a reporter construct harboring lacZ under the control of the R. etli virE promoter, we showed that the signal phenolic molecule acetosyringone (AS) induced R...
January 9, 2017: Journal of Bacteriology
https://www.readbyqxmd.com/read/28068385/the-arginine-pairs-and-c-termini-of-the-sso7c4-from-sulfolobus-solfataricus-participate-in-binding-and-bending-dna
#3
Bo-Lin Lin, Chin-Yu Chen, Chun-Hsiang Huang, Tzu-Ping Ko, Cheng-Hung Chiang, Kuan-Fu Lin, Yuan-Chih Chang, Po-Yen Lin, Hui-Hsu Gavin Tsai, Andrew H-J Wang
The Sso7c4 from Sulfolobus solfataricus forms a dimer, which is believed to function as a chromosomal protein involved in genomic DNA compaction and gene regulation. Here, we present the crystal structure of wild-type Sso7c4 at a high resolution of 1.63 Å, showing that the two basic C-termini are disordered. Based on the fluorescence polarization (FP) binding assay, two arginine pairs, R11/R22' and R11'/R22, on the top surface participate in binding DNA. As shown in electron microscopy (EM) images, wild-type Sso7c4 compacts DNA through bridging and bending interactions, whereas the binding of C-terminally truncated proteins rigidifies and opens DNA molecules, and no compaction of the DNA occurs...
2017: PloS One
https://www.readbyqxmd.com/read/28064034/highly-efficient-transformation-system-for-malassezia-furfur-and-malassezia-pachydermatis-using-agrobacterium-tumefaciens-mediated-transformation
#4
A M Celis, A M Vos, S Triana, C A Medina, N Escobar, S Restrepo, H A B Wösten, H de Cock
Malassezia spp. are part of the normal human and animal mycobiota but are also associated with a variety of dermatological diseases. The absence of a transformation system hampered studies to reveal mechanisms underlying the switch from the non-pathogenic to pathogenic life style. Here we describe, a highly efficient Agrobacterium-mediated genetic transformation system for Malassezia furfur and M. pachydermatis. A binary T-DNA vector with the hygromycin B phosphotransferase (hpt) selection marker and the green fluorescent protein gene (gfp) was introduced in M...
January 5, 2017: Journal of Microbiological Methods
https://www.readbyqxmd.com/read/28060264/peptide-derived-method-to-transport-genes-and-proteins-across-cellular-and-organellar-barriers-in-plants
#5
Jo-Ann Chuah, Yoko Horii, Keiji Numata
The capacity to introduce exogenous proteins and express (or down-regulate) specific genes in plants provides a powerful tool for fundamental research as well as new applications in the field of plant biotechnology. Viable methods that currently exist for protein or gene transfer into plant cells, namely Agrobacterium and microprojectile bombardment, have disadvantages of low transformation frequency, limited host range, or a high cost of equipment and microcarriers. The following protocol outlines a simple and versatile method, which employs rationally-designed peptides as delivery agents for a variety of nucleic acid- and protein-based cargoes into plants...
December 16, 2016: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28059116/further-characterization-of-maize-chlorotic-mottle-virus-and-its-synergistic-interaction-with-sugarcane-mosaic-virus-in-maize
#6
Qiang Wang, Chao Zhang, Chunyan Wang, Yajuan Qian, Zhenghe Li, Jian Hong, Xueping Zhou
Maize chlorotic mottle virus (MCMV) was first reported in maize in China in 2009. In this study we further analyzed the epidemiology of MCMV and corn lethal necrosis disease (CLND) in China. We determined that CLND observed in China was caused by co-infection of MCMV and sugarcane mosaic virus (SCMV). Phylogenetic analysis using four full-length MCMV cDNA sequences obtained in this study and the available MCMV sequences retrieved from GenBank indicated that Chinese MCMV isolates were derived from the same source...
January 6, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28054641/quorum-quenching-limits-quorum-sensing-exploitation-by-signal-negative-invaders
#7
Mélanie Tannières, Julien Lang, Claudie Barnier, Jacqui A Shykoff, Denis Faure
Some bacteria produce and perceive quorum-sensing (QS) signals that coordinate several behaviours, including the costly processes that are exoenzyme production and plasmid transfer. In the case of plasmid transfer, the emergence of QS signal-altered invaders and their policing are poorly documented. In Agrobacterium tumefaciens, the virulence Ti-plasmid encodes both synthesis and sensing of QS-signals, which promote its transfer from a donor to a recipient cell. Here, we reported that QS-altered A. tumefaciens mutants arose during experimental evolution...
January 5, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28053648/harnessing-anthocyanin-rich-fruit-a-visible-reporter-for-tracing-virus-induced-gene-silencing-in-pepper-fruit
#8
Jihyun Kim, Minkyu Park, Eun Soo Jeong, Je Min Lee, Doil Choi
BACKGROUND: Virus-induced gene silencing (VIGS) has become a powerful tool for post-genomic technology in plant species. This is important, especially in select plants, such as the pepper plant, that are recalcitrant to Agrobacterium-mediated transformation. Although VIGS in plants has been widely employed as a powerful tool for functional genomics, scattering phenotypic effects by uneven gene silencing has been implemented in order to overcome challenges in experiments with fruit tissues...
2017: Plant Methods
https://www.readbyqxmd.com/read/28050829/vnd6-induced-xylem-cell-differentiation-in-arabidopsis-cell-cultures
#9
Yoshihisa Oda
In vitro xylem differentiation is a powerful technique that can be used to elucidate the process of xylem development that occurs deep inside plant tissues in nature. The experimental procedure described here is designed to induce metaxylem vessel differentiation at exceptionally high frequency and synchronicity using genetically engineered Arabidopsis cell suspensions. By triggering a transcriptional switch, over 80 % of the cells synchronously differentiate into xylem cells within 32 h of treatment with estradiol...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28039549/expression-of-talaromyces-thermophilus-lipase-gene-in-trichoderma-reesei-by-homologous-recombination-at-the-cbh1-locus
#10
Xu Zhang, Liming Xia
CBH1 (cellobiohydrolase) comprises the majority of secreted proteins by Trichoderma reesei. For expression of Talaromyces thermophilus lipase gene in T. reesei, a self-designed CBH1 promoter was applied to drive the lipase gene expression cassette which was bracketed by flanking sequences of cbh1 gene for homologous recombination. Protoplast and Agrobacterium-mediated plasmid transformations were performed and compared, resultantly, transformation mediated by Agrobacterium was overall proved to be more efficient...
December 30, 2016: Journal of Industrial Microbiology & Biotechnology
https://www.readbyqxmd.com/read/28038423/citric-acid-application-for-denitrification-process-support-in-biofilm-reactor
#11
Artur Mielcarek, Joanna Rodziewicz, Wojciech Janczukowicz, Dorota Dabrowska, Slawomir Ciesielski, Arthur Thornton, Joanna Struk-Sokołowska
The study demonstrated that citric acid, as an organic carbon source, can improve denitrification in Anaerobic Sequencing Batch Biofilm Reactor (AnSBBR). The consumption rate of the organic substrate and the denitrification rate were lower during the period of the reactor's acclimatization (cycles 1-60; 71.5 mgCOD L(-1) h(-1) and 17.81 mgN L(-1) h(-1), respectively) than under the steady state conditions (cycles 61-180; 143.8 mgCOD L(-1) h(-1) and 24.38 mgN L(-1) h(-1)). The biomass yield coefficient reached 0...
December 22, 2016: Chemosphere
https://www.readbyqxmd.com/read/28026997/multifaceted-roles-of-the-ras-guanine-nucleotide-exchange-factor-chrgf-in-development-pathogenesis-and-stress-responses-of-colletotrichum-higginsianum
#12
Qiongnan Gu, Meijuan Chen, Junbin Huang, Y D Wei, Tom Hsiang, Lu Zheng
The infection process of Colletotrichum higginsianum, which causes a disease of crucifers, involves several key steps: conidial germination, appressorial formation, appressorial penetration, and invasive growth in host tissues. In this study, the ChRgf gene encoding a Ras guanine-nucleotide exchange factor protein was identified by screening T-DNA insertion mutants generated from Agrobacterium tumefaciens-mediated transformation (AtMT) that were unable to cause disease on the host Arabidopsis thaliana. Targeted gene deletion of ChRgf resulted in a null mutant (ΔChrgf-42) with defects in vegetative growth, hyphal morphology and conidiation, and poor surface attachment and low germination on hydrophobic surfaces, but there were no apparent differences in appressorial turgor pressure between the wild-type and the mutant...
December 27, 2016: Phytopathology
https://www.readbyqxmd.com/read/28026825/transformation-of-medicinal-plants-using-agrobacterium-tumefaciens
#13
Katarzyna Bandurska, Agnieszka Berdowska, Małgorzata Król
For many years attempts are made to develop efficient methods for transformation of medicinal plants via Agrobacterium tumefaciens. It is a soil bacteria which possess a natural ability to infect plants in places of injures which results in arise of cancerous growths (crown gall). This is possible thanks a transfer of fragment of Ti plasmid into plant cells and stable integration with a plant genome. Efficiency of medicinal plant transformation depends on many factors for example: Agrobacterium strain, methods and procedures of transformation as well as on plant species, type and age of the explants and regeneration conditions...
December 20, 2016: Postȩpy Higieny i Medycyny Doświadczalnej
https://www.readbyqxmd.com/read/28018866/bacillus-sp-qsi-1-modulate-quorum-sensing-signals-reduce-aeromonas-hydrophila-level-and-alter-gut-microbial-community-structure-in-fish
#14
Shuxin Zhou, An Zhang, Hongping Yin, Weihua Chu
Quorum sensing (QS) is a cell density dependent process that enables bacteria to communicate with each other based on the production, secretion and sensing of the auto-inducer molecules and then subsequently regulate virulence associated gene expression. Interrupting quorum sensing may represent a novel alternative approach to combat bacterial pathogen. Several bacteria can produce quorum quenching (QQ) enzymes. However, the role of QQ bacteria in shaping the microbiota and the level of N-acyl-homoserine lactones (AHLs, a prevalent type of QS molecules) producing bacteria remains largely unknown...
2016: Frontiers in Cellular and Infection Microbiology
https://www.readbyqxmd.com/read/27999977/auxin-synthesis-gene-tms1-driven-by-tuber-specific-promoter-alters-hormonal-status-of-transgenic-potato-plants-and-their-responses-to-exogenous-phytohormones
#15
Oksana O Kolachevskaya, Lidiya I Sergeeva, Kristyna Floková, Irina A Getman, Sergey N Lomin, Valeriya V Alekseeva, Elena B Rukavtsova, Yaroslav I Buryanov, Georgy A Romanov
Ectopic auxin overproduction in transgenic potato leads to enhanced productivity accompanied with concerted and occasional changes in hormonal status, and causing altered response of transformants to exogenous auxin or cytokinin. Previously, we generated potato transformants expressing Agrobacterium-derived auxin synthesis gene tms1 driven by tuber-specific patatin gene promoter (B33-promoter). Here, we studied the endogenous hormonal status and the response to exogenous phytohormones in tms1 transformants cultured in vitro...
December 20, 2016: Plant Cell Reports
https://www.readbyqxmd.com/read/27997767/characterization-of-silencing-suppressor-p24-of-grapevine-leafroll-associated-virus-2
#16
Mingjun Li, Jiao Zhang, Ming Feng, Xianyou Wang, Chen Luo, Qi Wang, Yuqin Cheng
Grapevine leafroll-associated virus 2 (GLRaV-2) p24 has been reported as an RNA-silencing suppressor (RSS). However, the mechanisms underlying p24's suppression of RNA silencing are unknown. Using Agrobacterium infiltration-mediated RNA-silencing assays, we showed that GLRaV-2 p24 is a strong RSS triggered by positive-sense GFP RNA, and that silencing suppression by p24 effectively blocks accumulation of small interfering RNAs. Deletion analyses showed that the region of amino acids 1-188, which contains all predicted α-helixes and β-strands, is required for p24's RSS activity...
December 20, 2016: Molecular Plant Pathology
https://www.readbyqxmd.com/read/27997398/toxicity-and-biofilm-based-selection-for-methyl-tert-butyl-ether-bioremediation-technology
#17
I M Guisado, J Purswani, L Catón-Alcubierre, J González-López, C Pozo
Extractive membrane biofilm reactor (EMBFR) technology offers productive solutions for volatile and semi-volatile compound removal from water bodies. In this study, the bacterial strains Paenibacillus etheri SH7(T) (CECT 8558), Agrobacterium sp. MS2 (CECT 8557) and Rhodococcus ruber strains A5 (CECT 8556), EE6 (CECT 8612) and EE1 (CECT 8555), previously isolated from fuel-contaminated sites, were tested for adherence on tubular semipermeable membranes in laboratory-scale systems designed for methyl tert-butyl ether (MTBE) bioremediation...
December 2016: Water Science and Technology: a Journal of the International Association on Water Pollution Research
https://www.readbyqxmd.com/read/27995281/the-use-of-stable-and-unstable-green-fluorescent-proteins-for-studies-in-two-bacterial-models-agrobacterium-tumefaciens-and-xanthomonas-campestris-pv-campestris
#18
Pilar Sabuquillo, Adela Gea, Isabel M Matas, Cayo Ramos, Jaime Cubero
Fluorescent proteins have been used to track plant pathogens to understand their host interactions. To be useful, the transgenic pathogens must present similar behaviour than the wild-type isolates. Herein, a GFP marker was used to transform two plant pathogenic bacteria, Agrobacterium and Xanthomonas, to localize and track the bacteria during infection. The transgenic bacteria were evaluated to determine whether they showed the same fitness than the wild-type strains or whether the expression of the GFP protein interfered in the bacterial activity...
December 19, 2016: Archives of Microbiology
https://www.readbyqxmd.com/read/27992646/divalent-cations-increase-dna-repair-activities-of-bacterial-6-4-photolyases
#19
Hongju Ma, Fan Zhang, Elisabeth Ignatz, Martin Suehnel, Peng Xue, Patrick Scheerer, LarsOliver Essen, Norbert Krauß, Tilman Lamparter
The (6-4) photolyases of the FeS-BCP group can be considered as the most ancient type among the large family of cryptochrome and photolyase flavoproteins. In contrast to other photolyases they contain an Fe-S cluster of unknown function, a DMRL chromophore, an interdomain loop which could interact with DNA, and a long C-terminal extension. We compared DNA repair and photoreduction of two members of the FeS-BCP family, Agrobacterium fabrum PhrB and Rhodobacter sphaeroides RsCryB, with a eukaryotic (6-4) photolyase from Ostreococcus, OsCPF, and a member of the class III CPD photolyases, PhrA from A...
December 19, 2016: Photochemistry and Photobiology
https://www.readbyqxmd.com/read/27992645/crystal-structures-of-bacterial-6-4-photolyase-mutants-with-impaired-dna-repair-activity
#20
Fan Zhang, Hongju Ma, Kalinga Bowatte, Dennis Kwiatkowski, Esther Mittmann, Heba Qasem, Norbert Krauss, Xiaoli Zeng, Zhong Ren, Patrick Scheerer, Xiaojing Yang, Tilman Lamparter
PhrB from Agrobacterium fabrum is the first prokaryotic photolyase which repairs (6-4) UV DNA-photoproducts. The protein harbors three cofactors, the enzymatically active FAD chromophore, a second chromophore, 6,7-dimethyl-8-ribityllumazine (DMRL) and a cubane-type Fe-S cluster. Tyr424 of PhrB is part of the DNA binding site and could provide an electron link to the Fe-S cluster. The PhrBY424F mutant showed reduced binding of lesion DNA and loss of DNA repair. The mutant PhrBI51W , is characterized by the loss of the DMRL chromophore, reduced photoreduction and reduced DNA repair capacity...
December 19, 2016: Photochemistry and Photobiology
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