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  1. Article ; Online: The Application of the Gibson Assembly Method in the Production of Two pKLS3 Vector-Derived Infectious Clones of Foot-and-Mouth Disease Virus

    Ploypailin Semkum / Nattarat Thangthamniyom / Penpitcha Chankeeree / Challika Keawborisuth / Sirin Theerawatanasirikul / Porntippa Lekcharoensuk

    Vaccines, Vol 11, Iss 1111, p

    2023  Volume 1111

    Abstract: The construction of a full-length infectious clone, essential for molecular virological study and vaccine development, is quite a challenge for viruses with long genomes or possessing complex nucleotide sequence structures. Herein, we have constructed ... ...

    Abstract The construction of a full-length infectious clone, essential for molecular virological study and vaccine development, is quite a challenge for viruses with long genomes or possessing complex nucleotide sequence structures. Herein, we have constructed infectious clones of foot-and-mouth disease virus (FMDV) types O and A by joining each viral coding region with our pKLS3 vector in a single isothermal reaction using Gibson Assembly (GA). pKLS3 is a 4.3-kb FMDV minigenome. To achieve optimal conditions for the DNA joining, each FMDV coding sequence was divided into two overlapping fragments of approximately 3.8 and 3.2 kb, respectively. Both DNA fragments contain the introduced linker sequences for assembly with the linearized pKLS3 vector. FMDV infectious clones were produced upon directly transfecting the GA reaction into baby hamster kidney-21 (BHK-21) cells. After passing in BHK-21 cells, both rescued FMDVs (rO189 and rNP05) demonstrated growth kinetics and antigenicity similar to their parental viruses. Thus far, this is the first report on GA-derived, full-length infectious FMDV cDNA clones. This simple DNA assembly method and the FMDV minigenome would facilitate the construction of FMDV infectious clones and enable genetic manipulation for FMDV research and custom-made FMDV vaccine production.
    Keywords Gibson assembly ; foot-and-mouth disease virus ; reverse genetics ; vaccine ; infectious clone ; Medicine ; R
    Subject code 630
    Language English
    Publishing date 2023-06-01T00:00:00Z
    Publisher MDPI AG
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  2. Article ; Online: Andrographolide and Deoxyandrographolide Inhibit Protease and IFN-Antagonist Activities of Foot-and-Mouth Disease Virus 3C pro

    Sirin Theerawatanasirikul / Varanya Lueangaramkul / Nattarat Thangthamniyom / Penpitcha Chankeeree / Ploypailin Semkum / Porntippa Lekcharoensuk

    Animals, Vol 12, Iss 15, p

    2022  Volume 1995

    Abstract: Foot-and mouth-disease (FMD) caused by the FMD virus (FMDV) is highly contagious and negatively affects livestock worldwide. The control of the disease requires a combination of measures, including vaccination; however, there is no specific treatment ... ...

    Abstract Foot-and mouth-disease (FMD) caused by the FMD virus (FMDV) is highly contagious and negatively affects livestock worldwide. The control of the disease requires a combination of measures, including vaccination; however, there is no specific treatment available. Several studies have shown that plant-derived products with antiviral properties were effective on viral diseases. Herein, antiviral activities of andrographolide (AGL), deoxyandrographolide (DAG), and neoandrographolide (NEO) against FMDV serotype A were investigated using an in vitro cell-based assay. The results showed that AGL and DAG inhibited FMDV in BHK-21 cells. The inhibitory effects of AGL and DAG were evaluated by RT-qPCR and exhibited EC50 values of 52.18 ± 0.01 µM (SI = 2.23) and 36.47 ± 0.07 µM (SI = 9.22), respectively. The intracellular protease assay revealed that AGL and DAG inhibited FMDV 3C pro with IC50 of 67.43 ± 0.81 and 25.58 ± 1.41 µM, respectively. Additionally, AGL and DAG significantly interfered with interferon (IFN) antagonist activity of the 3C pro by derepressing interferon-stimulating gene (ISGs) expression. The molecular docking confirmed that the andrographolides preferentially interacted with the 3C pro active site. However, NEO had no antiviral effect in any of the assays. Conclusively, AGL and DAG inhibited FMDV serotype A by interacting with the 3C pro and hindered its protease and IFN antagonist activities.
    Keywords 3C protease (3C pro ) ; andrographolide ; antiviral activity ; deoxyandrographolide ; diterpenoids ; foot-and-mouth disease virus ; Veterinary medicine ; SF600-1100 ; Zoology ; QL1-991
    Subject code 333
    Language English
    Publishing date 2022-08-01T00:00:00Z
    Publisher MDPI AG
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  3. Article: Molecular characterization of bovine ephemeral fever virus in Thailand between 2013 and 2017

    Chaisirirat, Thanawat / Pradit Sangthong / Pipat Arunvipas / Nantawan Petcharat / Nattarat Thangthamniyom / Wilairat Chumsing / Porntippa Lekcharoensuk

    Veterinary microbiology. 2018 Dec., v. 227

    2018  

    Abstract: Bovine ephemeral fever (BEF) is an arthropod-borne disease caused by bovine ephemeral fever virus (BEFV), a negative sense, single-stranded RNA virus. BEFV is endemic in tropical and sub-tropical regions including Thailand, a country in mainland ... ...

    Abstract Bovine ephemeral fever (BEF) is an arthropod-borne disease caused by bovine ephemeral fever virus (BEFV), a negative sense, single-stranded RNA virus. BEFV is endemic in tropical and sub-tropical regions including Thailand, a country in mainland Southeast Asia. However, there are few studies on BEFV and no available information regarding molecular characteristics of BEFV in Thailand. Therefore, the aims of this study were to genetically characterize Thai BEFVs and reveal their evolutions by phylogenetic analysis of G gene ectodomain sequences. From 2013 to 2017, blood samples were collected from bovine that matched with BEF case definition from three regions of Thailand. Thai BEFV G genes and a whole genome of an isolate, East Asia/TH/LRI0045/2016, were sequenced and characterized. Additionally, their phylogenies were constructed. This is the first report on genetics of BEFV in Southeast Asia. G ectodomain encoding region of Thai BEFV found during 2013–2017 are closely related to the second and third sub-clades of East Asia lineage. In addition, we observed mutation in the putative P’ ORF of all Thai BEFVs which generated a premature stop codon. Thai G gene sequences are closely related to those of mainland Chinese and Taiwanese isolates. The whole genomic sequences of Thai BEFV and East Asia/China/JT02 L/2002 possess common characteristics, suggesting shared evolutionary relationship between East and Southeast Asian strains. Further studies on relationship between animal translocation, circulation of BEFV in Greater Mekong subregion and acquisition of more G gene sequences may improve understanding of BEFV epidemiology in mainland Southeast Asia.
    Keywords Bovine ephemeral fever virus ; blood sampling ; bovine ephemeral fever ; cattle ; epidemiology ; mutation ; open reading frames ; phylogeny ; stop codon ; subtropics ; Thailand
    Language English
    Dates of publication 2018-12
    Size p. 1-7.
    Publishing place Elsevier B.V.
    Document type Article
    ZDB-ID 753154-0
    ISSN 1873-2542 ; 0378-1135
    ISSN (online) 1873-2542
    ISSN 0378-1135
    DOI 10.1016/j.vetmic.2018.10.013
    Database NAL-Catalogue (AGRICOLA)

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  4. Article: Development of quantitative competitive PCR for the detection of porcine circovirus type 2 DNA in pigs affected with PMWS

    Nattarat Thangthamniyom(Kasetsart University, Bangkok (Thailand). Faculty of Veterinary Medicine. Department of Microbiology and Immunology) E-mail:j2gole@yahoo.com / Porntippa Lekcharoensuk(Kasetsart University, Bangkok (Thailand). Faculty of Veterinary Medicine. Department of Microbiology and Immunology) / Tippawan Jantafong(Kasetsart University, Bangkok (Thailand). Faculty of Veterinary Medicine. Department of Microbiology and Immunology) / Nantawan Phecharat(Kasetsart University, Bangkok (Thailand). Faculty of Veterinary Medicine. Department of Microbiology and Immunology)

    Abstract: Porcine circovirus type 2 (PCV2) is an essential infectious agent of postweaning multisystemic wasting syndrome (PMWS). In this study, a quantitative competitive PCR (QC-PCR) assay was developed for monitoring PCV DNA in fecal swab from pigs in PMWS- ... ...

    Abstract Porcine circovirus type 2 (PCV2) is an essential infectious agent of postweaning multisystemic wasting syndrome (PMWS). In this study, a quantitative competitive PCR (QC-PCR) assay was developed for monitoring PCV DNA in fecal swab from pigs in PMWS-negative and -affected farms. The QC-PCR was based on competitive co-amplification of a 345 bp of the PCV type 2 with a known concentration of the competitor DNA, which produced a 513 bp fragment. The result demonstrated that the PCV2 DNA content obtained form the samples from the PMWS-affected farms higher than 1 pg/ul whereas those from the PMWS-negative farms was lower than 1 fg/ul with the exception of 10pg/ul of the sample F39. According to the results, the amount of PCV2 DNA load from clinical PMWS free and clinical PMWS affected farms were different significantly and this indicating that QC-PCR may be a useful tool for predicting of the occurrence of PMWS in pig farms.
    Keywords Swine ; Animal diseases ; Morbidity ; PCR ; DNA
    Language Thai
    Document type Article
    Database AGRIS - International Information System for the Agricultural Sciences and Technology

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