Journal of Microbiology

Journal Articles

Effects of Phosphorus‑dissolving Dark Septate Endophytes on the Growth of Blueberry: Qixin Luo , Rui Hou , Xiaojing Shang , Si Li; J. Microbiol. 2023;61(9):837-851. Published online October 5, 2023; DOI: https://doi.org/10.1007/s12275-023-00080-2

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Dark septate endophytes (DSEs) are widely distributed and improve plant growth. DSEs secrete large amounts of enzymes to mineralize insoluble phosphorus in soil and convert it into soluble phosphorus, promoting plant uptake of phosphorus. However, the effects of DSEs with phosphate-solubilizing ability on host plants need further study. In this study, phosphorusdissolving DSEs were screened for growth-promoting effects. We isolated, identified and characterized three DSE species (Thozetella neonivea, Pezicula ericae and Hyaloscyphaceae sp.) showing phosphate-solubilizing ability. The impact of single, dual or triple inoculation of DSEs on blueberry plant characteristics was studied. Their effects on colonization intensity, seedling biomass, nutrients in plants and soil, and activities of plant resistance enzymes and soil enzymes were markedly upregulated relative to the control (P < 0.05). The available phosphorus and acid phosphatase levels in different combinations were significantly increased. These findings indicate that the application of the three DSEs may be valuable in facilitating the cultivation of blueberry with a higher biomass and improved plant quality.

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Diversity and Functional Roles of Root-Associated Endophytic Fungi in Two Dominant Pioneer Trees Reclaimed from a Metal Mine Slag Heap in Southwest China
Bo Bi, Yuqing Xiao, Xiaonan Xu, Qianqian Chen, Haiyan Li, Zhiwei Zhao, Tao Li
Microorganisms.2024; 12(10): 2067. CrossRef
Short-term organic fertilizer substitution increases sorghum yield by improving soil physicochemical characteristics and regulating microbial community structure
Mengen Nie, Guangqian Yue, Lei Wang, Yizhong Zhang
Frontiers in Plant Science.2024;[Epub] CrossRef

Transposon insertion site sequencing (TIS) of Pseudomonas aeruginosa: Hongbaek Cho; J. Microbiol. 2021;59(12):1067-1074. Published online December 4, 2021; DOI: https://doi.org/10.1007/s12275-021-1565-y

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Abstract

Transposon insertion site sequencing (TIS) is a technique that determines the insertion profile of a transposon mutant library by massive parallel sequencing of transposon-genomic DNA junctions. Because the transposon insertion profile reflects the abundance of each mutant in the library, it provides information to assess the fitness contribution of each genetic locus of a bacterial genome in a specific growth condition or strain background. Although introduced only about a dozen years ago, TIS has become an important tool in bacterial genetics that provides clues to study biological functions and regulatory mechanisms. Here, I describe a protocol for generating high density transposon insertion mutant libraries and preparing Illumina sequencing samples for mapping the transposon junctions of the transposon mutant libraries using Pseudomonas aeruginosa as an example.

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Optimizing phage-based mutant recovery and minimizing heat effect in the construction of transposon libraries in Staphylococcus aureus
Sally W. Yousief, Nader Abdelmalek, Bianca Paglietti
Scientific Reports.2024;[Epub] CrossRef
The biological essence of synthetic lethality: Bringing new opportunities for cancer therapy
Meiyi Ge, Jian Luo, Yi Wu, Guobo Shen, Xi Kuang
MedComm – Oncology.2024;[Epub] CrossRef
Optimization of Transposon Mutagenesis Methods in Pseudomonas antarctica
Sangha Kim, Changhan Lee
Microorganisms.2023; 11(1): 118. CrossRef
Construction of high-density transposon mutant library of Staphylococcus aureus using bacteriophage ϕ11
Wonsik Lee
Journal of Microbiology.2022; 60(12): 1123. CrossRef

[Protocol]Construction of a multicopy genomic DNA library and its application for suppression analysis: Hongbaek Cho; J. Microbiol. 2019;57(12):1041-1047. Published online November 22, 2019; DOI: https://doi.org/10.1007/s12275-019-9417-8

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Abstract

Suppression analysis is used for the identification of new genes and genetic interactions when there is a notable phenotype available for genetic selection or screening. A random genomic DNA library constructed on a multi-copy plasmid is a useful tool for suppression analysis when one expects that an overdose of a few genes will suppress the phenotype. These libraries have been successfully used to determine the function of a gene by revealing genes whose functions are related to the gene of interest. They have also been used to identify the targets of chemical or biological agents by increasing the number of unaffected target gene products in a cell. In this article, I will discuss important considerations for constructing multicopy genomic DNA libraries. The protocol provided in this paper should be a useful guide for constructing genomic DNA libraries in many bacterial species for which multi-copy plasmids are available.

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Identification and characterization of the lipoprotein N -acyltransferase in Bacteroides
Krista M. Armbruster, Jiawen Jiang, Mariana G. Sartorio, Nichollas E. Scott, Jenna M. Peterson, Jonathan Z. Sexton, Mario F. Feldman, Nicole M. Koropatkin
Proceedings of the National Academy of Sciences.2024;[Epub] CrossRef

Review

MINIREVIEW] Modern and Simple Construction of Plasmid: Saving Time and Cost: Hideki Nakayama , Nobuo Shimamoto; J. Microbiol. 2014;52(11):891-897. Published online October 31, 2014; DOI: https://doi.org/10.1007/s12275-014-4501-6

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Abstract

Construction of plasmids has been occupying a significant fraction of laboratory work in most fields of experimental biology. Tremendous effort was made to improve the traditional method for constructing plasmids, in which DNA fragments digested with restriction enzymes were ligated. However, the traditional method remained to be a standard protocol more than 40 years. At last, several recent inventions are rapidly and completely replacing the traditional method, because they are far quicker with less cost, and requiring less material. We here introduce three such methods that cover up most of the cases. Moreover, they are complementary with each other. Our lab protocols are provided for “no strain, no pain” construction of plasmids.

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Tumor biology experimental course design based on the integration of molecular biology and metabolomics
Xinliang Zhu, Ting Tang
Cogent Education.2024;[Epub] CrossRef
Truncation-Free Genetic Code Expansion with Tetrazine Amino Acids for Quantitative Protein Ligations
Alex J. Eddins, Riley M. Bednar, Subhashis Jana, Abigail H. Pung, Lea Mbengi, Kyle Meyer, John J. Perona, Richard B. Cooley, P. Andrew Karplus, Ryan A. Mehl
Bioconjugate Chemistry.2023; 34(12): 2243. CrossRef
Involvement of GcvB small RNA in intrinsic resistance to multiple aminoglycoside antibiotics in Escherichia coli
Akira Muto, Simon Goto, Daisuke Kurita, Chisato Ushida, Hyota Himeno
The Journal of Biochemistry.2021; 169(4): 485. CrossRef
Flagellum-mediated motility in Pelotomaculum thermopropionicum SI
Tomoyuki Kosaka, Mutsumi Goda, Manami Inoue, Toshiharu Yakushi, Mamoru Yamada
Bioscience, Biotechnology, and Biochemistry.2019; 83(7): 1362. CrossRef
Natural killer cells unleashed: Checkpoint receptor blockade and BiKE/TriKE utilization in NK-mediated anti-tumor immunotherapy
Zachary B. Davis, Daniel A. Vallera, Jeffrey S. Miller, Martin Felices
Seminars in Immunology.2017; 31: 64. CrossRef
In vivo Assembly in Escherichia coli of Transformation Vectors for Plastid Genome Engineering
Yuyong Wu, Lili You, Shengchun Li, Meiqi Ma, Mengting Wu, Lixin Ma, Ralph Bock, Ling Chang, Jiang Zhang
Frontiers in Plant Science.2017;[Epub] CrossRef
Role of 100S ribosomes in bacterial decay period
Ksenia Shcherbakova, Hideki Nakayama, Nobuo Shimamoto
Genes to Cells.2015; 20(10): 789. CrossRef

Journal Article

NOTE] Comparison of the Genetic Structures Surrounding qnrA1 in Korean Enterobacter cloacae and Chinese Escherichia coli Strains Isolated in the Early 2000s: Evidence for qnrA Mobilization via Inc HI2 Type Plasmid: Sang Hoon Han , Young Ah Kim , Minggui Wang , Yangsoon Lee , Hae-Sun Chung , Jong Hwa Yum , Dongeun Yong , Kyungwon Lee , June Myung Kim; J. Microbiol. 2012;50(1):166-169. Published online February 27, 2012; DOI: https://doi.org/10.1007/s12275-012-1350-z

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Abstract: The flanking genetic structure of qnrA1 in Korean Enterobacter cloacae was identical to that of the Chinese Escherichia coli strain, the first qnrA1-carrying strain reported in Asia. Analysis of restriction enzyme sites and Southern blot hybridization results showed that qnrA1 was transferred between E. cloacae and E. coli via Inc HI2 type plasmid.

Research Support, Non-U.S. Gov'ts

NOTE] Complete Sequence and Organization of the Sphingobium chungbukense DJ77 pSY2 Plasmid: Sun-Mi Yeon , Young-Chang Kim; J. Microbiol. 2011;49(4):684-688. Published online September 2, 2011; DOI: https://doi.org/10.1007/s12275-011-1262-3

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Abstract: Sphingobium chungbukense DJ77 is capable of metabolizing priority chemicals of human health concern such as polycyclic aromatic hydrocarbons (PAHs), extracellular polysaccharide (EPS), and antibiotics. Here, we report the complete DNA and genetic organization of the plasmid pSY2 from strain DJ77. A DNA sequence analysis revealed that pSY2 comprises 18,779 bp encoding 22 open reading frames (ORFs) with 59.5% G+C content. The ORFs on pSY2 were classified into DNA replication, conjugative function, transposition, plasmid stability/partition, and other functional groups (transport, fatty acid biosynthesis, stress, and growth rate regulation). Three ORFs on pSY2 were hypothetical proteins.

Molecular Analysis of a Prolonged Spread of Klebsiella pneumoniae Co-producing DHA-1 and SHV-12 β-Lactamases: Young Kyung Yoon , Hye Won Cheong , Hyunjoo Pai , Kyoung Ho Roh , Jeong Yeon Kim , Dae Won Park , Jang Wook Sohn , Seung Eun Lee , Byung Chul Chun , Hee Sun Sim , Min Ja Kim; J. Microbiol. 2011;49(3):363-368. Published online June 30, 2011; DOI: https://doi.org/10.1007/s12275-011-0491-9

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Abstract: The study investigated molecular mechanisms for prolonged nosocomial spread of multidrug-resistant Klebsiella pneumoniae co-producing plasmid-mediated AmpC β-lactamase DHA-1 and extended-spectrum β-lactamase SHV-12. Forty-eight clinical isolates of K. pneumonia, resistant to the extended-spectrum cephalosporins, were collected in a 750-bed university hospital over a year. The isolates were characterized for PCR-based β-lactamase genotypes, isoelectric focusing and pulsed-field gel electrophoresis (PFGE) profiles. Resistance transfer was performed by plasmid conjugation and confirmed by a duplex-PCR and Southern hybridization. On β-lactamase typing, the strains producing only the DHA-1 enzyme (n=17) or co-producing DHA-1 and SHV-12 enzymes (n=15) were predominant. Judging from a one year-distribution of PFGE profiles, the co-producer was spread primarily with single clonal expansion of the PFGE-type A with subtypes (n=14), whereas the strains producing only DHA-1 enzyme were spread simultaneously with the PFGE-type A (n=11) and other PFGE types (n=6). Transconjugants of the co-producers were confirmed to harbor either both blaDHA-1 and blaSHV-12 or only the blaDHA-1. In conclusion, this study indicated that the persistent nosocomial spread of multidrug-resistant K. pneumoniae strains was primarily associated with expansion of a clone harboring both the blaDHA-1 and blaSHV-12 or the blaDHA-1 only, and to a lesser extent with the horizontal transfer of the resistant plasmids. Our observations have clinical implication for the control and prevention of nosocomial dissemination of multidrug-resistant K. pneumoniae strains.

Characterization of Plasmid pSY3 in Sphingobium chungbukense DJ77: Sun-Mi Yeon , Young-Chang Kim; J. Microbiol. 2009;47(6):796-800. Published online February 4, 2010; DOI: https://doi.org/10.1007/s12275-009-0329-x

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Abstract: This study determined the complete nucleotide sequence of the plasmid pSY3 from Sphingobium chungbukense DJ77. It was 35,735 bp long with a G+C content of 61.9%. Forty open reading frames (ORFs) were found. We predicted these ORFs would encode proteins associated with plasmid replication, conjugative transfer, transposition of genes, plasmid stability/partition, hypothetical protein, and some other functions. Genes for biodegradation were not found. No other plasmid homologous to pSY3 in the overall nucleotide sequence or gene organization could be found in the NCBI database.

Cloning and Molecular Characterization of a Novel Rolling-Circle Replicating Plasmid, pK1S-1, from Bacillus thuringiensis subsp. kurstaki K1: Ming Shun Li , Jong Yul Roh , Xueying Tao , Zi Niu Yu , Zi Duo Liu , Qin Liu , Hong Guang Xu , Hee Jin Shim , Yang-Su Kim , Yong Wang , Jae Young Choi , Yeon Ho Je; J. Microbiol. 2009;47(4):466-472. Published online September 9, 2009; DOI: https://doi.org/10.1007/s12275-009-0020-2

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Abstract: Bacillus thuringiensis, an entomopathogenic bacterium belonging to the B. cereus group, harbors numerous extra-chromosomal DNA molecules whose sizes range from 2 to 250 kb. In this study, we used a plasmid capture system (PCS) to clone three small plasmids from B. thuringiensis subsp. kurstaki K1 which were not found in B. thuringiensis subsp. kurstaki HD-1, and determined the complete nucleotide sequence of plasmid pK1S-1 (5.5 kb). Of the six putative open reading frames (ORF2-ORF7) in pK1S-1, ORF2 (MobK1) showed approximately 90% aa identity with the Mob-proteins of pGI2 and pTX14-2, which are rolling circle replicating group VII (RCR group VII) plasmids from B. thuringiensis. In addition, a putative origin of transfer (oriT) showed 95.8% identity with those of pGI2 and pTX14-2. ORF3 (RepK1) showed relatively low aa identity (17.8~25.2%) with the Rep protein coded by RCR plasmids, however. The putative double- strand origin of replication (dso) and single-strand origin of replication (sso) of pK1S-1 exhibited approximately 70% and 64% identities with those of pGI2 and pTX14-2. ORF6 and 7 showed greater than 50% similarities with alkaline serine protease, which belongs to the subtilase family. The other 2 ORFs were identified as hypothetical proteins. To determine the replicon of pK1S-1, seven subclones were contructed in the B. thuringiensis ori-negative pHT1K vector and were electroporated into a plasmid cured B. thuringiensis strain. The 1.6 kb region that included the putative ORF3 (Rep1K), dso and ORF4, exhibited replication ability. These findings identified pK1S-1 as a new RCR group VII plasmid, and determined its replication region.

Journal Article

Mobilization Functions of the Bacteriocinogenic Plasmid pRJ6 of Staphylococcus aureus: Marcus Livio Varella Coelho , Hilana Ceotto , Danielle Jannuzzi Madureira , Ingolf F. Nes , Maria do Carmo de Freire Bastos; J. Microbiol. 2009;47(3):327-336. Published online June 26, 2009; DOI: https://doi.org/10.1007/s12275-009-0044-7

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Abstract: Plasmid pRJ6 is the first known bacteriocinogenic mobilizable (Mob) plasmid of Staphylococcus aureus. Its Mob region is composed of four mob genes (mobCDAB) arranged as an operon, a genetic organization uncommon among S. aureus Mob plasmids. oriTpRJ6 was detected in a region of 431 bp, positioned immediately upstream of mobC. This region, when cloned into pCN37, was able to confer mobilization to the recombinant plasmid only in the presence of pRJ6. The entire Mob region, including oriTpRJ6, is much more similar to Mob regions from several coagulase-negative staphylococci plasmids, although some remarkable similarities with S. aureus Mob plasmids can also be noted. These similarities include the presence within oriTpRJ6 of the three mcb (MobC binding sites), firstly described in pC221 and pC223, an identical nick site also found in these same plasmids, and a nearly identical srapC223 site (sequence recognized by MobA). pRJ6 was successfully transferred to S. epidermidis by conjugation in the presence of the conjugative plasmid pGO1. Altogether these findings suggest that pRJ6 might have been originally a coagulase-negative staphylococci plasmid that had been transferred successfully to S. aureus.

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Class 1 and Class 2 Integrons and Plasmid-Mediated Antibiotic Resistance in Coliforms Isolated from Ten Rivers in Northern Turkey: Osman Birol Ozgumus , Cemal Sandalli , Ali Sevim , Elif Celik-Sevim , Nuket Sivri; J. Microbiol. 2009;47(1):19-27. Published online February 20, 2009; DOI: https://doi.org/10.1007/s12275-008-0206-z

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Abstract: We aimed to determine the molecular mechanisms of antibiotic resistance in coliforms isolated from ten rivers in northern region of Turkey. A total of 183 isolates were tested for antimicrobial susceptibility by disk diffusion and agar dilution methods. Resistance to ampicillin, streptomycin, trimethoprim, tetracycline, and chloramphenicol was detected in 58%, 51.9%, 24%, 28.4%, and 12.5%, respectively. Twelve (6.5%) phylogenetically distant organisms were detected to harbor self-transmissible plasmids ranging 52 to >147 kb in sizes. Resistances to ampicillin, tetracycline, trimethoprim, streptomycin, and nalidixic acid were commonly transferable traits. Transferable nalidixic acid-resistant strains harbored qnrS gene, which was the first report of plasmid-mediated quinolone resistance in bacteria of environmental origin in Turkey. Fourteen and five coliforms harbored class 1 and class 2 integrons, respectively, and some of them were located on transferable plasmids. Sequence analyses of variable regions of the class 1 and 2 integrons harbored various gene cassettes, dfrA1, dfr2d, dfrA7, dfrA16, dfrA17, aadA1, aadA5, blaOXA-30, and sat1. A gene cassette array, dfrA16 has been demonstrated for the first time in a Citrobacter koseri isolate. Class 1 and class 2-bearing strains were clustered in different groups by BOX-PCR fingerprinting. Rivers in the northern Turkey may act as receptacle for the multi-drug resistant enterobacteria and can serve as reservoirs of the antimicrobial resistance determinants in the environment. The actual risk to public health is the transfer of resistance genes from the environmental bacteria to human pathogens.

Prevalence of Tetracycline Resistance Genes in Greek Seawater Habitats: Theodora L. Nikolakopoulou , Eleni P. Giannoutsou , Adamandia A. Karabatsou , Amalia D. Karagouni; J. Microbiol. 2008;46(6):633-640. Published online December 24, 2008; DOI: https://doi.org/10.1007/s12275-008-0080-8

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Abstract: The presence of selected tetracycline resistance (TcR) genes was studied in different Greek seawater habitats, originated from wastewater treatment facilities, fishfarm, and coastal environments. The methods employed included assessment of the presence of twelve gene clusters by PCR, followed by hybridization with specific probes, in habitat extracted DNA, TcR bacteria, and exogenous isolated plasmids conferring TcR. The direct DNA-based analysis showed that tet(Α) and tet(K) genes were detected in all habitats, whilst tet(C) and tet(E) were present in fishfarm and wastewater effluent samples and tet(M) was detected in fishfarm and coastal samples. Resistance genes tet(A), tet(C), tet(K), and tet(M) were detected in 60 of the 89 isolates screened. These isolates were identified by fatty acid methyl ester analysis (FAME) as Stenotrophomonas, Acinetobacter, Pseudomonas, Bacillus, and Staphylococcus strains. The presence of the TcR genes in 15% of the bacterial isolates coincided with the presence of IncP plasmids. A habitat-specific dissemination of IncP alpha plasmids in wastewater effluent isolates and of IncP beta plasmids in fishfarm isolates was observed. Exogenous isolation demonstrated the presence of plasmids harbouring TcR genes in all the habitats tested. Plasmids were shown to carry tet(A), tet(C), tet(E), and tet(K) genes. It is concluded that TcR genes are widespread in the seawater habitats studied and often occur on broad host range plasmids that seem to be well disseminated in the bacterial communities.

pVC, a Small Cryptic Plasmid from the Environmental Isolate of Vibrio cholerae MP-1: Ruifu Zhang , Yanling Wang , Pak Chow Leung , Ji-Dong Gu; J. Microbiol. 2007;45(3):193-198.; DOI: https://doi.org/2543 [pii]

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Abstract: A marine bacterium was isolated from Mai Po Nature Reserve of Hong Kong and identified as Vibrio cholerae MP-1. It contains a small plasmid designated as pVC of 3.8 kb. Four open reading frames (ORFs) are identified on the plasmid, but none of them shows homology to any known protein. Database search indicated that a 440 bp fragment is 96% identical to a fragment found in a small plasmid of another V. cholerae. Further experiments demonstrated that a 2.3 kb EcoRI fragment containing the complete ORF1, partial ORF4 and their intergenic region could self-replicate. Additional analyses revealed that sequence upstream of ORF1 showed the features characteristic of theta type replicons. Protein encoded by ORF1 has two characteristic motifs existed in most replication initiator proteins (Rep): the leucine zipper (LZ) motif located at the N-terminal region and the alpha helix-turn-alpha helix motif (HTH) located at the C-terminal end. The results suggest that pVC replicates via the theta type mechanism and is likely a novel type of theta replicon.

Evidence of Indigenous NAH Plasmid of Naphthalene Degrading Pseudomonas putida PpG7 Strain Implicated in Limonin Degradation: Moushumi Ghosh , Abhijit Ganguli , Meenakshi Mallik; J. Microbiol. 2006;44(5):473-479.; DOI: https://doi.org/2451 [pii]

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Abstract: A well characterized naphthalene-degrading strain, Pseudomonas putida PpG7 was observed to utilize limonin, a highly-oxygenated triterpenoid compound as a sole source of carbon and energy. Limonin concentrations evidenced a 64% reduction over 48 h of growth in batch cultures. Attempts were made to acquire a plasmid-less derivative via various methods (viz. Ethidium Bromide, SDS, elevated temperature & mitomycin C), among which the method involving mitomycin C (20 μg/ml) proved successful. Concomitant with the loss of plasmid in P. putida PpG7 strain, the cured derivative was identified as a lim- phenotype. The lim+ phenotype could be conjugally transferred to the cured derivative. Based on the results of curing with mitomycin C, conjugation studies and presence of ndo gene encoding naphthalene 1,2 dioxygenase, it was demonstrated that genes for the limonin utilization were encoded on an 83 kb indigenous transmissible Inc. P9 NAH plasmid in Pseudomonas putida PpG7 strain.

Isolation and Characterization of a Rhodococcus Species Strain Able to Grow on ortho- and para-Xylene: Jung Yeon Jang , Dockyu Kim , Hyun Won Bae , Ki Young Choi , Jong-Chan Chae , Gerben J. Zylstra , Young Min Kim , Eungbin Kim; J. Microbiol. 2005;43(4):325-330.; DOI: https://doi.org/2258 [pii]

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Abstract: Rhodococcus sp. strain YU6 was isolated from soil for the ability to grow on o-xylene as the sole carbon and energy source. Unlike most other o-xylene-degrading bacteria, YU6 is able to grow on p-xylene. Numerous growth substrate range experiments, in addition to the ring-cleavage enzyme assay data, suggest that YU6 initially metabolizes o- and p-xylene by direct aromatic ring oxidation. This leads to the formation of dimethylcatechols, which was further degraded largely through meta-cleavage pathway. The gene encoding meta-cleavage dioxygenase enzyme was PCR cloned from genomic YU6 DNA using previously known gene sequence data from the o-xylene-degrading Rhodococcus sp. strain DK17. Subsequent sequencing of the 918-bp PCR product revealed a 98% identity to the gene, encoding methylcatechol 2,3-dioxygenase from DK17. PFGE analysis followed by Southern hybridization with the catechol 2,3-dioxygenase gene demonstrated that the gene is located on an approximately 560-kb megaplasmid, designated pJYJ1 <br>

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