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  1. Article: Cloning and production of nucleoprotein of swine influenza virus in E.coli

    Areerat Pangpeng(Kasetsart University, Bangkok (Thailand). Faculty of Veterinary Medicine. Department of Microbiology and Immunology) E-mail:areejum@hotmail.com / Nantawan Phecharat(Kasetsart University, Bangkok (Thailand). Faculty of Veterinary Medicine. Department of Microbiology and Immunology) / Porntippa Lekcharoensuk(Kasetsart University, Bangkok (Thailand). Faculty of Veterinary Medicine. Department of Microbiology and Immunology)

    Abstract: Influenza A virus is a causative agent of swine influenza which is an acute respiratory disease affecting pigs at all ages leading to considerable economic losses of swine industries. Monitoring and surveillance of swine influenza virus (SIV) infection ... ...

    Abstract Influenza A virus is a causative agent of swine influenza which is an acute respiratory disease affecting pigs at all ages leading to considerable economic losses of swine industries. Monitoring and surveillance of swine influenza virus (SIV) infection and herd immunity are essential for disease prevention and control. However, it requires an effective diagnostic tool such as ELISA. Nucleoprotein (NP) is a major group specific and conserved protein suitable to be use as antigen for the detection of all subtypes of influenza A viruses. The objective of this study was to production and purification of the recombinant NP protein. Total RNA was extracted from swine influenza virus (A/swine/IA/1930) suspension. cDNA was synthesis from the total RNA and NP gene was amplified using NP specific primers which designed to contain KpnI and SalI at 5 ends of the forward and reverse primers, respectively. The NP gene was ligated to the vector, pQE80L, prior to be transformed into E.coli strain DH5 alpha. In E.coli. The expressed protein reached the highest yield when the expression was induced using 0.2 mM IPTG at 37 deg C for 24 hours. Majority of the protein was insoluble and accumulated as inclusion bodies. The insoluble NP protein was dissolved in 8 M Urea pH 8.0 and purified using a nickel-charged resin-affinity column. The protein was then neutralized using stepwise dilution method. The recombinant protein had a molecular weight approximately 56 kDa and reacted strongly to convalescent serum from a pig infected with SIV. The results showed that the recombinant NP protein was successfully produced. It is immunogenic and ready for further diagnostic application
    Keywords Swine ; Viruses ; Animal diseases ; Viroses ; Influenzavirus ; Molecular cloning ; Genetic engineering ; Animal production ; Nucleoproteins
    Language Thai
    Document type Article
    Database AGRIS - International Information System for the Agricultural Sciences and Technology

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  2. 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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    Inter-library loan at ZB MED

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