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Publication Metadata only Limited cross-reactivity of mouse monoclonal antibodies against Dengue virus capsid protein among four serotypes(2012-11-22) Megumi Noda; Promsin Masrinoul; Chaweewan Punkum; Chonlatip Pipattanaboon; Pongrama Ramasoota; Chayanee Setthapramote; Tadahiro Sasaki; Mikiko Sasayama; Akifumi Yamashita; Takeshi Kurosu; Kazuyoshi Ikuta; Tamaki Okabayashi; Mahidol University; Tohoku University; Osaka UniversityFlaviviridae. We prepared monoclonal antibodies against DENV capsid protein from mice immunized with DENV-2 and determined the cross-reactivity with each serotype of DENV and Japanese encephalitis virus. Methods and results: To clarify the relationship between... stained the nuclei of infected cells and were found to be specific to the capsid protein. Three were specific to DENV-2, while the other two were cross-reactive with DENV-2 and DENV-4. No monoclonal antibodies were cross-reactive with all four serotypesPublication Metadata only Shrimp laminin receptor binds with capsid proteins of two additional shrimp RNA viruses YHV and IMNV(2011-07-01) Nattaphon Busayarat; Saengchan Senapin; Moltira Tonganunt; Kornsunee Phiwsaiya; Watcharachai Meemetta; Sasimanas Unajak; Sarawut Jitrapakdee; Chu Fang Lo; Amornrat Phongdara; Prince of Songkla University; Thailand National Center for Genetic Engineering and Biotechnology; Mahidol University; National Taiwan UniversityLaminin receptor (Lamr) in shrimp was previously proposed to be a potential receptor protein for Taura syndrome virus (TSV) based on yeast two-hybrid assays. Since shrimp Lamr bound to the VP1 capsid protein of TSV, we were interested to know... whether capsid/envelope proteins from other shrimp viruses would also bind to Lamr. Thus, capsid/envelope encoding genes from 5 additional shrimp viruses were examined. These were Penaeus stylirostris densovirus (PstDNV), white spot syndrome virus (WSSVPublication Metadata only Production and characterization of anti-dengue capsid antibodies suggesting the N terminus region covering the first 20 amino acids of dengue virus capsid protein is predominantly immunogenic in mice(2009-06-30) Chunya Puttikhunt; Prapapun Ong-ajchaowlerd; Tanapan Prommool; Sutha Sangiambut; Janjuree Netsawang; Thawornchai Limjindaporn; Prida Malasit; Watchara Kasinrerk; Faculty of Medicine, Siriraj Hospital, Mahidol University; Mahidol University; Chiang Mai University; Armed Forces Research Institute of Medical Sciences, ThailandWe produced monoclonal and polyclonal antibodies to the capsid (C) protein of dengue serotype 2 virus (DV2 C). First, a maltose-binding protein fused to DV2 C protein (MBP-C) was overproduced in E. coli. The affinity-purified MBP-C protein... was cleaved by factor Xa protease to obtain a recombinant DV2 C protein, which was then used for mouse immunizations. Two hybridoma cell lines producing anti-C Mabs as well as anti-C polyclonal antibody were successfully generated and characterizedPublication Metadata only Interaction study of a novel Macrobrachium rosenbergii effector caspase with B2 and capsid proteins of M.rosenbergii nodavirus reveals their roles in apoptosis(2015-08-01) Supak Youngcharoen; Saengchan Senapin; Tareerat Lertwimol; Siwaporn Longyant; Paisarn Sithigorngul; Timothy W. Flegel; Parin Chaivisuthangkura; Srinakharinwirot University; Mahidol University; Thailand National Center for Genetic Engineering and Biotechnology; Thailand National Metal and Materials Technology Centerwith caspase 3 of shrimp. The functions of the Mrcasp, B2 and capsid proteins of M. rosenbergii nodavirus (. MrNV) were assayed in Sf-9 cells. The results showed that Mrcasp induce apoptotic morphology cells; however, capsid protein of MrNV could inhibit... apoptotic cells whereas B2 could neither induce nor inhibit apoptotic cells by DAPI staining. The protein interaction between Mrcasp and viral MrNV structure revealed that Mrcasp did not bind to B2 or capsid protein whereas B2 and capsid proteins could bindPublication Metadata only Production of monoclonal antibodies specific to Macrobrachium rosenbergii nodavirus using recombinant capsid protein(2012-03-20) Pradit Wangman; Saengchan Senapin; Parin Chaivisuthangkura; Siwaporn Longyant; Sombat Rukpratanporn; Paisarn Sithigorngul; Srinakharinwirot University; Mahidol University; Thailand National Center for Genetic Engineering and Biotechnology; Chulalongkorn UniversityThe gene encoding the capsid protein of Macrobrachium rosenbergii nodavirus (MrNV) was cloned into pGEX-6P-1 expression vector and then transformed into the Escherichia coli strain BL21. After induction, capsid protein-glutathione-S- transferase... (GST-MrNV; 64 kDa) was produced. The recombinant protein was separated using SDS-PAGE, excised from the gel, electro-eluted and then used for immunization for monoclonal antibody (MAb) production. Four MAbs specific to the capsid protein were selectedPublication Metadata only Co-expression of double-stranded RNA and viral capsid protein in the novel engineered Escherichia coli DualX-B15(DE3) strain(2021-12-01) Kitti Wuthisathid; Thawatchai Chaijarasphong; Charoonroj Chotwiwatthanakun; Monsicha Somrit; Kallaya Sritunyalucksana; Ornchuma Itsathitphaisarn; Mahidol University; Thailand National Center for Genetic Engineering and Biotechnologyconstructed a novel E. coli strain that is able to co-express viral capsid proteins and dsRNA in the same E. coli cell. Results: A novel bacterial strain DualX-B15(DE3) was engineered to be both protease- and RNase III-deficiency via P1 phage transduction.... The results revealed that it could simultaneously express recombinant proteins and dsRNA. Conclusion: Co-expression of viral capsid proteins and dsRNA in the same cell has been shown to be feasible. Not only could this platform serve as a basis for future costPublication Metadata only Highly conserved nucleotide sequence and its deduced amino acids of the 5′-noncoding region and the capsid protein of a Bangkok isolate dengue-3 virus(2000-12-01) W. H. Attatippaholkun; M. K. Attatippaholkun; A. Nisalak; D. W. Vaughn; B. L. Innis; Mahidol University; Thailand Institute of Scientific and Technological Research (TISTR); Armed Forces Research Institute of Medical Sciences, Thailand; Walter Reed Army Institute of Research- 3′. The nucleotide sequence of the 5′-noncoding region and the capsid protein of dengue-3virus (a Bangkok isolate; CH53489 isolated by USAMC-AFRIMS in 1973) has been analyzed in both forward and reverse directions. The PCR-based cycle sequencing... and arginine. The homology of the nucleotide sequence between the two dengue-3 virus revealed 97%. The deduced amino acid sequences from the nucleotides 95-465 of the two viruses showed the same indicating highly conserved capsid proteins. Multiple alignmentPublication Metadata only Sustained replication of dengue pseudoinfectious virus lacking the capsid gene by trans-complementation in capsid-producing mosquito cells(2013-06-01) Sutha Sangiambut; Amporn Suphatrakul; Rungtawan Sriburi; Poonsook Keelapang; Chunya Puttikhunt; Watchara Kasinrerk; Prida Malasit; Nopporn Sittisombut; Thailand National Center for Genetic Engineering and Biotechnology; Chiang Mai University; Mahidol Universityof the dengue virus capsid protein served as host cells for the trans-complementation of partially capsid-deleted viral RNA generated in vitro. Transfection of the partially capsid-deleted viral RNA into the anchored capsid-expressing C6/36 cells resulted... in moderate titers of infectious virus. Progeny viruses multiplied in the capsid trans-complementing C6/36 cells for up to three weeks, but only initiated single rounds of replication in Vero cells lacking the capsid protein. Employing this transPublication Metadata only Binding of shrimp cellular proteins to Taura syndrome viral capsid proteins VP1, VP2 and VP3(2006-12-01) Saengchan Senapin; Amornrat Phongdara; Thailand National Center for Genetic Engineering and Biotechnology; Mahidol University; Prince of Songkla Universitysystem was employed to study interactions between three Taura syndrome viral capsid proteins (VP1-VP3) and proteins from a cDNA library of the black tiger shrimp Penaeus monodon. VP1 interacted with β-actin, elongation factor 1α (EF1α), lysozyme (Lys...) and laminin receptor/ribosomal protein p40 (Lamr/p40) containing a putative palindromic laminin binding region LMWWML. VP2 interacted with β-actin and EF1α, while VP3 bound to the same proteins as VP1 except for Lamr/p40. In vitro pull-down assays confirmedPublication Metadata only The effect of capsid mutations on HIV-1 uncoating(2007-02-05) Pathawut Wacharapornin; Darat Lauhakirti; Prasert Auewarakul; Mahidol UniversityEfficient uncoating requires not only an optimal cellular environment, but also some intrinsic properties of the viral capsid protein itself. Using an in vitro uncoating model, we demonstrated that substitution of each serine residue with alanine... at the three major phosphorylation sites of HIV-1 capsid protein, i.e. Ser-109, Ser-149 and Ser-178, could significantly reduce uncoating activity of purified core particles. We also showed that the core stability of mutant viruses was lower than