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Li Xu, Di-Si Lin, Jing Yang, Jian Li, Bei Li
OBJECTIVE: To construct the KbvR gene of LuxR family deletion mutant and complementation strains from Klebsiella pneumoniae NTUH-K2044 and analyze the effect of KbvR on bacterial growth, biofilm formation and capsular synthesis. METHODS: A KbvR gene deletion mutant strain was constructed using the suicide vector pKO3-Km, and the gene fragment including KbvR coding region, promoter area and transcription termination area were amplified and cloned into pGEM-T-easy plasmid to construct KbvR complementation strain...
October 20, 2016: Nan Fang Yi Ke da Xue Xue Bao, Journal of Southern Medical University
Tzu-Lung Lin, Yi-Jiun Pan, Pei-Fang Hsieh, Chun-Ru Hsu, Meng-Chuan Wu, Jin-Town Wang
Analysis of the genome of Klebsiella pneumoniae NTUH-K2044 strain revealed the presence of two clustered regularly interspaced short palindromic repeats (CRISPR) arrays separated with CRISPR-associated (cas) genes. Carbapenem-resistant K. pneumoniae isolates were observed to be less likely to have CRISPR-Cas than sensitive strains (5/85 vs. 22/132). Removal of the transcriptional repressor, H-NS, was shown to prevent the transformation of plasmids carrying a spacer and putative proto-spacer adjacent motif (PAM)...
2016: Scientific Reports
Kuo-Feng Hua, Feng-Ling Yang, Hsiao-Wen Chiu, Ju-Ching Chou, Wei-Chih Dong, Chien-Nan Lin, Chai-Yi Lin, Jin-Town Wang, Lan-Hui Li, Huan-Wen Chiu, Yi-Chich Chiu, Shih-Hsiung Wu
Klebsiella pneumoniae (strain 43816, K2 serotype) induces interleukin-1β (IL-1β) secretion, but neither the bacterial factor triggering the activation of these inflammasome-dependent responses nor whether they are mediated by NLRP3 or NLRC4 is known. In this study, we identified a capsular polysaccharide (K1-CPS) in K. pneumoniae (NTUH-K2044, K1 serotype), isolated from a primary pyogenic liver abscess (PLA K. pneumoniae), as the Klebsiella factor that induces IL-1β secretion in an NLRP3-, ASC-, and caspase-1-dependent manner in macrophages...
September 2015: Infection and Immunity
Harald Eriksson, Barbara Maciejewska, Agnieszka Latka, Grazyna Majkowska-Skrobek, Marios Hellstrand, Öjar Melefors, Jin-Town Wang, Andrew M Kropinski, Zuzanna Drulis-Kawa, Anders S Nilsson
Klebsiella pneumoniae phages vB_KpnP_SU503 (SU503) and vB_KpnP_SU552A (SU552A) are virulent viruses belonging to the Autographivirinae subfamily of Podoviridae that infect and kill multi-resistant K. pneumoniae isolates. Phages SU503 and SU552A show high pairwise nucleotide identity to Klebsiella phages KP34 (NC_013649), F19 (NC_023567) and NTUH-K2044-K1-1 (NC_025418). Bioinformatic analysis of these phage genomes show high conservation of gene arrangement and gene content, conserved catalytically active residues of their RNA polymerase, a common and specific lysis cassette, and form a joint cluster in phylogenetic analysis of their conserved genes...
April 2015: Viruses
Tzu-Lung Lin, Yi-Ping Chuang, Yu-Tsung Huang, Pei-Fang Hsieh, Yi-Tsung Lin, Jin-Town Wang
BACKGROUND: Klebsiella pneumoniae has emerged worldwide as a cause of pyogenic liver abscess (PLA) often complicated by meningitis and endophthalmitis. Early detection of this infectious disease will improve its clinical outcome. Therefore, we tried to isolate immunodominant proteins secreted by K. pneumoniae strains causing PLA. RESULTS: The secreted proteins of the NTUH-K2044 strain were separated by two-dimensional electrophoresis and then immunoblotted using convalescent sera from patients with K...
2014: BMC Microbiology
Ramy A Fodah, Jacob B Scott, Hok-Hei Tam, Pearlly Yan, Tia L Pfeffer, Ralf Bundschuh, Jonathan M Warawa
Klebsiella pneumoniae is a bacterial pathogen of worldwide importance and a significant contributor to multiple disease presentations associated with both nosocomial and community acquired disease. ATCC 43816 is a well-studied K. pneumoniae strain which is capable of causing an acute respiratory disease in surrogate animal models. In this study, we performed sequencing of the ATCC 43816 genome to support future efforts characterizing genetic elements required for disease. Furthermore, we performed comparative genetic analyses to the previously sequenced genomes from NTUH-K2044 and MGH 78578 to gain an understanding of the conservation of known virulence determinants amongst the three strains...
2014: PloS One
Tzu-Lung Lin, Pei-Fang Hsieh, Yu-Tsung Huang, Wei-Ching Lee, Yi-Ting Tsai, Po-An Su, Yi-Jiun Pan, Chun-Ru Hsu, Meng-Chuan Wu, Jin-Town Wang
BACKGROUND: Klebsiella pneumoniae causing community-acquired pyogenic liver abscess complicated with metastatic meningitis and endophthalmitis has emerged recently, most frequently associated with the K1 capsular type. METHODS: A bacteriophage (NTUH-K2044-K1-1) that infects K. pneumoniae NTUH-K2044 (capsular type K1) was isolated and characterized. RESULTS: The phage infected all K1 strains, and none of the strains with other capsular types...
December 1, 2014: Journal of Infectious Diseases
Pablo Ivan Pereira Ramos, Renata Christina Picão, Luiz Gonzaga Paula de Almeida, Nicholas Costa B Lima, Raquel Girardello, Ana Carolina P Vivan, Danilo E Xavier, Fernando G Barcellos, Marsileni Pelisson, Eliana C Vespero, Claudine Médigue, Ana Tereza Ribeiro de Vasconcelos, Ana Cristina Gales, Marisa Fabiana Nicolás
BACKGROUND: Klebsiella pneumoniae is an important opportunistic pathogen associated with nosocomial and community-acquired infections. A wide repertoire of virulence and antimicrobial resistance genes is present in K. pneumoniae genomes, which can constitute extra challenges in the treatment of infections caused by some strains. K. pneumoniae Kp13 is a multidrug-resistant strain responsible for causing a large nosocomial outbreak in a teaching hospital located in Southern Brazil. Kp13 produces K...
2014: BMC Genomics
Kai Wei Kelvin Lee, Krithika Arumugam, Rikky Wenang Purbojati, Qi Xiang Martin Tay, Rohan Benjamin Hugh Williams, Staffan Kjelleberg, Scott A Rice
Klebsiella pneumoniae is ubiquitous in the environment and is a member of a three-species biofilm model. We compared the genome sequence of an environmental isolate, K. pneumoniae strain KP-1, to those of two clinical strains (NTUH-K2044 and MGH 78578). KP-1 possesses strain-specific prophage sequences that distinguish it from the clinical strains.
2013: Genome Announcements
M H Zulkifli, L K Teh, L S Lee, Z A Zakaria, M Z Salleh
Klebsiella pneumoniae PR04 was isolated from a patient hospitalized in Malaysia. The draft genome sequence of K. pneumoniae PR04 shows differences compared to the reference sequences of K. pneumoniae strains MGH 78578 and NTUH-K2044 in terms of their genomic structures.
2013: Genome Announcements
Vijaya Bharathi Srinivasan, Govindan Rajamohan
Klebsiella pneumoniae has been frequently associated with nosocomial infections. Efflux systems are ubiquitous transporters that also function in drug resistance. Genome analysis of K. pneumoniae strain NTUH-K2044 revealed the presence of ∼15 putative drug efflux systems. We discuss here for the first time the characterization of a putative SMR-type efflux pump, an ebrAB homolog (denoted here as kpnEF) with respect to Klebsiella physiology and the multidrug-resistant phenotype. Analysis of hypermucoviscosity revealed direct involvement of kpnEF in capsule synthesis...
September 2013: Antimicrobial Agents and Chemotherapy
Vijaya Bharathi Srinivasan, Amitabha Mondal, Manjunath Venkataramaiah, Neeraj Kumar Chauhan, Govindan Rajamohan
Klebsiella pneumoniae is a Gram-negative bacillus that causes serious infections in immunocompromised human hosts and exhibits significant multidrug resistance. In this study, we identified a novel lysR-family regulator (designated oxyR(KP)) in the genome of K. pneumoniae NTUH-K2044 whose functions have remained enigmatic so far. Functional characterization of the putative lysR regulator oxyR(KP) with respect to cellular physiology and antimicrobial susceptibility was performed by generating an isogenic mutant, ΔoxyR(KP) in a hypervirulent clinical isolate of K...
July 2013: Microbiology
Tzu-Lung Lin, Feng-Ling Yang, An-Suei Yang, Hung-Pin Peng, Tsung-Lin Li, Ming-Daw Tsai, Shih-Hsiung Wu, Jin-Town Wang
Mucoviscosity-associated gene A (magA) of Klebsiella pneumoniae contributes to K1 capsular polysaccharide (CPS) biosynthesis. Based on sequence homology and gene alignment, the magA gene has been predicted to encode a Wzy-type CPS polymerase. Sequence alignment with the Wzy_C and RfaL protein families (which catalyze CPS or lipopolysaccharide (LPS) biosynthesis) and topological analysis has suggested that eight highly conserved residues, including G308, G310, G334, G337, R290, P305, H323, and N324, were located in a hypothetical loop region...
2012: PloS One
Soojin Lee, Borim Kim, Minkyu Oh, Youngrok Kim, Jinwon Lee
Meso-secondary alcohol dehydrogenases (meso-SADH) from Klebsiella oxytoca KCTC1686 and Klebsiella pneumoniae KCTC2242 were codon optimized and expressed in Escherichia coli W3110. The published gene data of K. pneumoniae NTUH-K2044 (NCBI accession number AP006725), K. pneumoniae 342 (NCBI accession number CP000964), and K. pneumoniae MGH 78578 (NCBI accession number CP000647), were compared with the meso-SADH sequences of each strain, respectively. Codon-optimized meso-SADH enzymes of K. oxytoca and K. pneumoniae showed approximately twofold to fivefold increased enzyme activities for acetoin reduction over native enzymes...
July 2013: Bioprocess and Biosystems Engineering
Vijaya Bharathi Srinivasan, Manjunath Venkataramaiah, Amitabha Mondal, Vasanth Vaidyanathan, Tanvi Govil, Govindan Rajamohan
BACKGROUND: The diffusion of antibiotics through the outer membrane is primarily affected by the porin super family, changes contribute to antibiotic resistance. Recently we demonstrated that the CpxAR two-component signaling system alters the expression of an uncharacterized porin OmpC(KP), to mediate antimicrobial resistance in K. pneumoniae. PRINCIPAL FINDINGS: In this study, functional characterization of the putative porin OmpC(KP) (denoted kpnO) with respect to antimicrobial susceptibility and virulence was evaluated by generating an isogenic mutant, ΔkpnO in a clinical isolate of K...
2012: PloS One
Diana P Cruz, Mónica G Huertas, Marcela Lozano, Lina Zárate, María Mercedes Zambrano
BACKGROUND: Klebsiella pneumoniae can be found in environmental habitats as well as in hospital settings where it is commonly associated with nosocomial infections. One of the factors that contribute to virulence is its capacity to form biofilms on diverse biotic and abiotic surfaces. The second messenger Bis-(3'-5')-cyclic dimeric GMP (c-di-GMP) is a ubiquitous signal in bacteria that controls biofilm formation as well as several other cellular processes. The cellular levels of this messenger are controlled by c-di-GMP synthesis and degradation catalyzed by diguanylate cyclase (DGC) and phophodiesterase (PDE) enzymes, respectively...
2012: BMC Microbiology
Vijaya Bharathi Srinivasan, Vasanth Vaidyanathan, Amitabha Mondal, Govindan Rajamohan
BACKGROUND: Klebsiella pneumoniae is a gram-negative, non-motile, facultative anaerobe belonging to the Enterobacteriaceae family of the γ-Proteobacteria class in the phylum Proteobacteria. Multidrug resistant K. pneumoniae have caused major therapeutic problems worldwide due to emergence of extended-spectrum β-lactamase producing strains. Two-component systems serve as a basic stimulus-response coupling mechanism to allow organisms to sense and respond to changes in many different environmental conditions including antibiotic stress...
2012: PloS One
Pei-Fang Hsieh, Tzu-Lung Lin, Feng-Ling Yang, Meng-Chuan Wu, Yi-Jiun Pan, Shih-Hsiung Wu, Jin-Town Wang
Klebsiella pneumoniae is the common cause of a global emerging infectious disease, community-acquired pyogenic liver abscess (PLA). Capsular polysaccharide (CPS) and lipopolysaccharide (LPS) are critical for this microorganism's ability to spread through the blood and to cause sepsis. While CPS type K1 is an important virulence factor in K. pneumoniae causing PLA, the role of LPS in PLA is not clear. Here, we characterize the role of LPS O antigen in the pathogenesis of K. pneumoniae causing PLA. NTUH-K2044 is a LPS O1 clinical strain; the presence of the O antigen was shown via the presence of 1,3-galactan in the LPS, and of sequences that align with the wb gene cluster, known to produce O-antigen...
2012: PloS One
Chun-Ru Hsu, Tzu-Lung Lin, You-Ci Chen, Huei-Chi Chou, Jin-Town Wang
Klebsiella pneumoniae community-acquired pyogenic liver abscess (PLA) is an emerging infectious disease. The rmpA gene (for regulator of mucoid phenotype A) has been reported to be associated with PLA in prevalence studies. NTUH-K2044, a K1 PLA isolate, carries three rmpA/A2 genes: two large-plasmid-carried genes (p-rmpA and p-rmpA2) and one chromosomal gene (c-rmpA). In this study, we re-examined the role of rmpA/A2 in PLA pathogenesis to clarify the relationship of rmpA/A2 and capsular serotype to virulence...
December 2011: Microbiology
Yi-Jiun Pan, Tzu-Lung Lin, Chun-Ru Hsu, Jin-Town Wang
Phagocytosis resistance is an important virulence factor in Klebsiella pneumoniae. Dictyostelium has been used to study the interaction between phagocytes and bacteria because of its similarity to mammalian macrophages. In this study, we used a Dictyostelium model to investigate genes for resistance to phagocytosis in NTUH-K2044, a strain of K. pneumoniae causing pyogenic liver abscess that is highly resistant to phagocytosis. A total of 2,500 transposon mutants were screened by plaque assay, and 29 of them permitted phagocytosis by Dictyostelium...
March 2011: Infection and Immunity
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