Journal of Microbiology

Journal Article

Identification and Characterization of HEPN‑MNT Type II TA System from Methanothermobacter thermautotrophicus ΔH: Wonho Choi , Anoth Maharjan , Hae Gang Im , Ji-Young Park , Jong-Tae Park , Jung-Ho Park; J. Microbiol. 2023;61(4):411-421. Published online April 18, 2023; DOI: https://doi.org/10.1007/s12275-023-00041-9

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Toxin-antitoxin (TA) systems are widespread in bacteria and archaea plasmids and genomes to regulate DNA replication, gene transcr!ption, or protein translation. Higher eukaryotic and prokaryotic nucleotide-binding (HEPN) and minimal nucleotidyltransferase (MNT) domains are prevalent in prokaryotic genomes and constitute TA pairs. However, three gene pairs (MTH304/305, 408/409, and 463/464) of Methanothermobacter thermautotropicus ΔH HEPN-MNT family have not been studied as TA systems. Among these candidates, our study characterizes the MTH463/MTH464 TA system. MTH463 expression inhibited Escherichia coli growth, whereas MTH464 did not and blocked MTH463 instead. Using site-directed MTH463 mutagenesis, we determined that amino acids R99G, H104A, and Y106A from the R[ɸX]4-6H motif are involved with MTH463 cell toxicity. Furthermore, we established that purified MTH463 could degrade MS2 phage RNA, whereas purified MTH464 neutralized MTH463 activity in vitro. Our results indicate that the endonuclease toxin MTH463 (encoding a HEPN domain) and its cognate antitoxin MTH464 (encoding the MNT domain) may act as a type II TA system in M. thermautotropicus ΔH. This study provides initial and essential information studying TA system functions, primarily archaea HEPN-MNT family.

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Extensive Genomic Rearrangement of Catalase-Less Cyanobloom-Forming Microcystis aeruginosa in Freshwater Ecosystems
Minkyung Kim, Jaejoon Jung, Wonjae Kim, Yerim Park, Che Ok Jeon, Woojun Park
Journal of Microbiology.2024; 62(11): 933. CrossRef

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

Development of a stringent ELISA protocol to evaluate anti-viral hemorrhagic septicemia virus-specific antibodies in olive flounder Paralichthys olivaceus with improved specificity: Hyoung Jun Kim , Jeong Su Park , Se Ryun Kwon; J. Microbiol. 2015;53(7):481-485. Published online June 27, 2015; DOI: https://doi.org/10.1007/s12275-015-5101-9

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Abstract

Olive flounder were vaccinated with polyinosinic:polycytidylic acid [Poly (I:C)] to prevent viral hemorrhagic septicemia (VHS). Vaccine efficacy was verified by detection of anti- VHS virus (VHSV) antibodies using enzyme-linked immunosorbent assay (ELISA). In the study, ELISA absorbance values of the negative control group [Poly (I:C)-MEM10] were saturated when an ELISA protocol, that includes pretreatment of the fish sera with 5% skim milk, was used. However, the saturated OD values in the negative control did not correlate with a specific immune response against VHSV, because the group showed low survival rate (only 10%) following the VHSV challenge. Also, OD values of Poly (I:C)- VHSV group were high, and the group showed high survival rate (97.5%) against VHSV challenge test. It was suggested that the high OD values were possibly due to the presence of anti-fetal bovine serum (FBS) cross-reactivity. To compensate this, we subtracted the absorbance of infectious hematopoietic necrosis (IHNV)-Ag plates from those of the VHSV-Ag plates. However, the average value for the Poly (I:C)-VHSV group (0.167) was lower than expected even though high survival rate. We used an advanced ELISA system to pre-treat fish sera with 5% skim milk and two novirhabdoviruses as capture antigens as well as 50% FBS. The corrected absorbance values for pre-treated fish sera from the negative control Poly (I:C)-MEM10 and experimental Poly (I:C)-VHSV groups averaged 0.033 and 0.579, respectively. The specific VHSV antibody response of the vaccinated group was assessed using fish sera pretreated with skim milk and FBS and by calculating the corrected absorbance values from ELISA with two novirhabdovirus capture antigens.

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Development of an indirect ELISA for detection of the adaptive immune response of black carp (Mylopharyngodon piceus)
Tongtong Wang, Shanshan Jin, Ruoxuan Lv, Yuting Meng, Guozhong Li, Yuxing Han, Qiusheng Zhang
Journal of Immunological Methods.2023; 521: 113550. CrossRef
Detection of Salmonid IgM Specific to the Piscine Orthoreovirus Outer Capsid Spike Protein Sigma 1 Using Lipid-Modified Antigens in a Bead-Based Antibody Detection Assay
Lena Hammerlund Teige, Subramani Kumar, Grethe M. Johansen, Øystein Wessel, Niccolò Vendramin, Morten Lund, Espen Rimstad, Preben Boysen, Maria K. Dahle
Frontiers in Immunology.2019;[Epub] CrossRef
Production of a monoclonal antibody against of muskellunge (Esox masquinongy) IgM heavy chain and its use in development of an indirect ELISA for titrating circulating antibodies against VHSV-IVB
Mohamed Faisal, Isaac F. Standish, Mary Ann Vogelbein, Elena V. Millard, Stephen L. Kaattari
Fish & Shellfish Immunology.2019; 88: 464. CrossRef
Phylogenetic analysis and duplex RT-PCR detection of viral hemorrhagic septicemia virus in olive flounder (Paralichthys olivaceus) from Korea
Jee Youn Hwang, Seongdo Lee, Thanthrige Thiunuwan Priyathilaka, Hyerim Yang, Hyukjae Kwon, Mun Gyeong Kwon, Seong Don Hwang, Myoung-Jin Kim, Jehee Lee
Aquaculture.2018; 484: 242. CrossRef
Herpesvirus Infection Induces both Specific and Heterologous Antiviral Antibodies in Carp
Julio M. Coll
Frontiers in Immunology.2018;[Epub] CrossRef
Validation of a serum neutralization test for detection of antibodies specific to cyprinid herpesvirus 3 in infected common and koi carp (Cyprinus carpio)
J Cabon, L Louboutin, J Castric, S Bergmann, G Bovo, M Matras, O Haenen, N J Olesen, T Morin
Journal of Fish Diseases.2017; 40(5): 687. CrossRef
The amino-terminal domain of ORF149 of koi herpesvirus is preferentially targeted by IgM from carp populations surviving infection
F. Torrent, A. Villena, P. A. Lee, W. Fuchs, S. M. Bergmann, J. M. Coll
Archives of Virology.2016; 161(10): 2653. CrossRef

Detection of Viruses in Farmed Rainbow Trout (Oncorhynchus mykiss) in Korea by RT-LAMP Assay: Rungkarn Suebsing , Jeong-Ho Kim , Seok Ryel Kim , Myung-Ae Park , Myung-Joo Oh; J. Microbiol. 2011;49(5):741-746. Published online November 9, 2011; DOI: https://doi.org/10.1007/s12275-011-1209-8

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Abstract: The viral diseases have been the serious problem in salmonid farming, and rainbow trout is not an exception. In this study, routine surveys were conducted for detecting of viruses in farmed rainbow trout (Oncorhynchus mykiss) in Korea during 2009-2010. Head kidneys from individual fish were employed for virus detection by using a reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay. Infectious pancreatic necrosis virus (IPNV), infectious hematopoietic necrosis virus (IHNV), and viral hemorrhagic septicemia virus (VHSV) were the target viruses in this study. 53.5% (46/86) were found to be IPNV-positive, while IHNV and VHSV showed RT-LAMP negative during examination for 2 years. Ten IPNV-positive samples were randomly selected for viral isolation and the cells showing CPEs were subjected to RT-LAMP, RT-PCR, and direct sequencing. Phylogenetic analysis showed that the rainbow trout isolate has high similarity homologies with the VR-299 strain, as previously described.

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