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Zhuang Yao, Xiaoming Liu, Jae Min Shim, Kang Wook Lee, Hyun-Jin Kim, Jeong Hwan Kim
Nine bacilli with fibrinolytic activities were isolated from Doenjang, a traditional Korean fermented soy food. Among them, RSB 34 showed the strongest activity and identified as Bacillus amyloliquefaciens by 16S rRNA and recA genes sequencing. During growth on LB up to 96 h, RSB34 showed the highest fibrinolytic activity (83.23 mU/μl) at 48 h. Three bands of 23, 27, and 42 kDa in size were observed when culture supernatant was analyzed by SDS-PAGE and 27 and 42 kDa bands by fibrin zymography. A gene encoding the 27 kDa major fibrinolytic enzyme, AprE34, was cloned by PCR...
October 25, 2016: Journal of Microbiology and Biotechnology
Jenni Kyyriäinen, Xavier Ekolle Ndode-Ekane, Asla Pitkänen
Platelet-derived growth factor receptor β (PDGFRβ) is upregulated after brain injury and its depletion results in the blood-brain barrier (BBB) damage. We investigated the time-window and localization of PDGFRβ expression in mice with intrahippocampal kainic acid-induced status epilepticus (SE) and in rats with lateral fluid-percussion-induced traumatic brain injury (TBI). Tissue immunohistochemistry was evaluated at several time-points after SE and TBI. The distribution of PDGFRβ was analyzed, and its cell type-specific expression was verified with double/triple-labeling of astrocytes (GFAP), NG2 cells, and endothelial cells (RECA-1)...
October 25, 2016: Glia
Zhe Wang, Fuwu Zhang, Zhantong Wang, Xiao Fu, Albert Jin, Bryant C Yung, Jing Fan, Xiangyu Yang, Gang Niu, Xiaoyuan Chen
Molecular design of biomaterials with unique features reca-pitulating nature's niche to influence biological activities has been a prolific area of investigation in chemistry and material science. The extracellular matrix (ECM) provides a wealth of bioactive molecules in supporting cell proliferation, migra-tion and differentiation. The well-patterned fibril and inter-twining architecture of the ECM profoundly influences cell behavior and development. Inspired by those features from the ECM, we attempted to integrate essential biological fac-tors from the ECM to design bioactive molecules to construct artificial self-supportive ECM mimics to advance stem cell culture...
October 24, 2016: Journal of the American Chemical Society
Nisha Tak, Esha Awasthi, Garima Bissa, Raju Ram Meghwal, Euan K James, Janet S Sprent, Hukam S Gehlot
Phylogenetically diverse Ensifer strains associated with five species of Tephrosia growing in alkaline soils of semi-arid regions of the Thar Desert were characterized using multi locus sequence analysis. Based on 16S rRNA and four protein-coding housekeeping gene (recA, atpD, glnII and dnaK) sequences, the Tephrosia-Ensifer strains were genetically different from the type strains of Ensifer saheli, Ensifer kostiensis, Ensifer terangae (African origin) and Ensifer psoraleae (Asiatic origin). One strain, Ensifer sp...
October 4, 2016: Systematic and Applied Microbiology
Oihane Irazoki, Jesús Aranda, Timo Zimmermann, Susana Campoy, Jordi Barbé
In addition to its role in DNA damage repair and recombination, the RecA protein, through its interaction with CheW, is involved in swarming motility, a form of flagella-dependent movement across surfaces. In order to better understand how SOS response modulates swarming, in this work the location of RecA and CheW proteins within the swarming cells has been studied by using super-resolution microscopy. Further, and after in silico docking studies, the specific RecA and CheW regions associated with the RecA-CheW interaction have also been confirmed by site-directed mutagenesis and immunoprecipitation techniques...
2016: Frontiers in Microbiology
Zhongqiu Chen, Wen Ling, Guangdong Shang
Pseudomonas putida KT2440 is a saprophytic, generally recognized as safe microorganism that plays important roles in the biodegradation and production of value-added chemicals. Chromosomal gene deletion of P. putida KT2440 usually involves time-consuming gene coning, conjugal transfer and counterselection. Recently, we developed a P. putida KT2440 markerless gene deletion method based on recombineering and Cre/loxP site-specific recombination. PCR-based λ Red recombineering circumvents the tedious cloning steps and is more amenable to high-throughput manipulation...
October 7, 2016: FEMS Microbiology Letters
Masaki Odahara, Yusuke Kobayashi, Toshiharu Shikanai, Yoshiki Nishimura
The chloroplast (cp) genome is organized as nucleoids that are dispersed throughout the cp stroma. Previously, a cp homolog of bacterial recombinase RecA (cpRECA) was shown to be involved in maintenance of cp genome integrity by repairing damaged cpDNA and by suppressing aberrant recombination between short dispersed repeats (SDRs) in the moss Physcomitrella patens. Here, overexpression and knockdown analysis of cpRECA in the green alga Chlamydomonas reinhardtii revealed that cpRECA was involved in cp nucleoid dynamics as well as having a role in maintaining cp genome integrity...
October 17, 2016: Plant Physiology
Xianchao Pan, Qiaoxia Zhang, Sujun Qu, Shuheng Huang, Huicong Wang, Hu Mei
ATP-binding cassette (ABC) exporters mediate vital transport of a variety of molecules across the lipid bilayer in all organisms. To explore the allosteric effect of ATP binding at the asymmetric ATPase sites, molecular dynamics simulations were performed on the nucleotide-binding domains (NBDs) of a heterodimeric exporter TM287/288 in 4 different ATP-bound states. The results showed that ATP bound at the degenerate site can maintain a semi-open conformation of NBD1-NBD2, which may be defective in ATP hydrolysis...
October 12, 2016: Integrative Biology: Quantitative Biosciences From Nano to Macro
Sundru Manjulata Devi, Subramanian Aishwarya, Prakash M Halami
The present study was aimed to evaluate the diversity and probiotic properties of Lactobacillus plantarum-group cultures from vegetable origin. First, genotypic diversity of L. plantarum (n=34) was achieved by PCR of Random Amplified Polymorphic DNA and recA gene-specific multiplex PCR. The isolates were segregated into five groups namely, Lactobacillus pentosus, Lactobacillus paraplantarum, Lactobacillus arizonensis, Lactobacillus plantarum subsp. plantarum and argentoratensis. Further discrimination was achieved by restriction fragment length polymorphism of probiotic adhesion genes viz...
October 5, 2016: Systematic and Applied Microbiology
María J Lorite, José David Flores-Félix, Álvaro Peix, Juan Sanjuán, Encarna Velázquez
In this study four Mesorhizobium strains isolated from Lotus corniculatus nodules in Granada (Spain) were characterized. Their 16S rRNA gene sequences were closely related to those of M. albiziae LMG 23507(T) and M. chacoense Pr5(T) showing 99.4 and 99.2% similarity values, respectively. The analysis of concatenated rpoB, recA, atpD and glnII genes showed they formed a cluster with internal similarities higher than 97%. The closest species also were M. albiziae LMG 23507(T) and M. chacoense Pr5(T) showing similarity values lower than 92% in rpoB, recA and glnII genes and lower than 96...
September 24, 2016: Systematic and Applied Microbiology
Paulo G Hofstatter, Alexander K Tice, Seungho Kang, Matthew W Brown, Daniel J G Lahr
Recombinase enzymes promote DNA repair by homologous recombination. The genes that encode them are ancestral to life, occurring in all known dominions: viruses, Eubacteria, Archaea and Eukaryota. Bacterial recombinases are also present in viruses and eukaryotic groups (supergroups), presumably via ancestral events of lateral gene transfer. The eukaryotic recA genes have two distinct origins (mitochondrial and plastidial), whose acquisition by eukaryotes was possible via primary (bacteria-eukaryote) and/or secondary (eukaryote-eukaryote) endosymbiotic gene transfers (EGTs)...
October 12, 2016: Proceedings. Biological Sciences
Rodrigo Rojas, Claudio D Miranda, Javier Santander, Jaime Romero
The VPAP30 strain was isolated as the highly predominant bacteria from an episode of massive larval mortality occurring in a commercial culture of the Chilean scallop Argopecten purpuratus. The main aims of this study were, to characterize and identify the pathogenic strain using biochemical and molecular methods to demonstrate its pathogenic activity on scallop larvae, to characterize its pathogenic properties and to describe the chronology of this pathology. The pathogenic strain was identified as Vibrio tubiashii based on its phenotypic properties and the sequence analysis of its 16S rRNA and housekeeping genes (ftsZ, gapA, gyrB, mreB, pyrH, recA, rpoA and topA)...
2016: Frontiers in Microbiology
James P R Connolly, Andrew J Roe
: We recently discovered that exposure of enterohaemorrhagic Escherichia coli (EHEC) to D-serine resulted in accumulation of this unusual amino acid, induction of the SOS regulon and downregulation of type III secretion that is essential for efficient colonization of the host. Here, we have investigated the physiological relevance of this elevated SOS response, which is of particular interest given the presence of Stx-toxin carrying lysogenic prophages on the EHEC chromosome that are activated during the SOS response...
October 3, 2016: Journal of Bacteriology
Marie Wrande, Helene Andrews-Polymenis, Donna J Twedt, Olivia Steele-Mortimer, Steffen Porwollik, Michael McClelland, Leigh A Knodler
Intestinal epithelial cells provide an important colonization niche for Salmonella enterica serovar Typhimurium (S Typhimurium) during gastrointestinal infections. In infected epithelial cells, a sub-population of S. Typhimurium damage their internalization vacuole, leading to escape from the Salmonella-containing vacuole (SCV) and extensive proliferation in the cytosol. Little is known about the bacterial determinants of nascent SCV lysis and subsequent survival and replication of Salmonella in the cytosol...
October 3, 2016: Infection and Immunity
Javier Dubert, Sabela Balboa, María Regueira, Adrián González-Castillo, Bruno Gómez-Gil, Jesús L Romalde
Seven isolates were obtained from different culture stages of carpet shell clam (Ruditapes decussatus) reared in a bivalve hatchery (Galicia, NW Spain). Three groups were differentiated by genotyping techniques and phenotypic profiles and representative trains were selected to further taxonomic studies. These strains were studied by a polyphasic approach and in basis of the phylogenetic analysis based on concatenated sequences of the five housekeeping genes ftsZ, gyrB, pyrH, recA and rpoA formed a tight group into the Mediterranei clade of the genus Vibrio...
September 20, 2016: Systematic and Applied Microbiology
Gangyi Chen, Juan Dong, Yi Yuan, Na Li, Xin Huang, Xin Cui, Zhuo Tang
Nucleic acid amplification is the core technology of molecular biology and genetic engineering. Various isothermal amplification techniques have been developed as alternatives to polymerase chain reaction (PCR). However, most of these methods can only detect single stranded nucleic acid. Herein, we put forward a simple solution for opening double-stranded DNA for isothermal detection methods. The strategy employs recombination protein from E. coli (RecA) to form nucleoprotein complex with single-stranded DNA, which could scan double-stranded template for homologous sites...
September 30, 2016: Scientific Reports
Siwei Shi, Huanhuan Chen, Hua Jiang, Yueqing Xie, Lei Zhang, Ninghuan Li, Chencen Zhu, Junsheng Chen, Han Luo, Jiaxian Wang, Lei Feng, Huili Lu, Jianwei Zhu
Soluble expression of recombinant therapeutic proteins in Escherichia coli (E. coli) has been a challenging task in biopharmaceutical development. In this study, a novel self-cleavable tag Zbasic-intein has been constructed for the soluble expression and purification of a recombinant cytokine, human interleukin-15 (IL-15). We screened several solubilizing tags fused with the self-cleavable Mycobacterium tuberculosis recA mini-intein ∆I-CM and demonstrated that Zbasic tag can significantly improve the solubility of the product with correspondent to the intein activity...
September 28, 2016: Applied Microbiology and Biotechnology
Cheng-Tai Huang, Chi-Te Liu, Shiang-Jiuun Chen, Wen-Yuan Kao
Crotalaria zanzibarica is an exotic and widely distributed leguminous plant in Taiwan. The relationship between C. zanzibarica and its rhizobial symbionts has been suggested to contribute to its successful invasion. A rhizobial strain (designed as CzR2) isolated from the root nodules of C. zanzibarica and cultivated in standard YEM medium displayed pleomorphism, with cells ranging between 2 and 10 μm in length and some branching. In the present study, we identified this rhizobial strain, investigated the causes of pleomorphism, and examined the nodules formed...
September 29, 2016: Microbes and Environments
Zhongyang Sun, Jia Deng, Haizhen Wu, Qiyao Wang, Yuanxing Zhang
Edwardsiella tarda is a Gram-negative pathogenic bacterium in aquaculture which can cause hemorrhagic septicemia in fish. Many secreted proteins have been identified as virulent factors of E. tarda. While the virulent phenotype is based on the expression regulation of the virulent genes. It is important to understand the expression profile of the virulent genes. Quantitative RT-PCR is one of the preferred methods for determining different gene expressions. It is important to select the most stable gene as a reference in E...
September 23, 2016: Journal of Microbiology and Biotechnology
Jie Wang, Qi Liu, Jia Feng, Jun-Ping Lv, Shu-Lian Xie
Severe eutrophication and harmful cyanobacterial blooms of freshwater ecosystems is a persistent environmental topic in recent decades. Pyrogallol (polyphenol) was confirmed to exhibit one of the most intensive inhibitory effects on the Microcystis aeruginosa. In this study, the expression of genes, release of microcystins (MCs) and antioxidant system of pyrogallol on Microcystis aeruginosa TY001 were investigated. The results revealed that the expression of stress response genes (prx, ftsH, grpE and fabZ) and DNA repair genes (recA and gyrB) were up-regulated...
September 16, 2016: Ecotoxicology and Environmental Safety
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