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HIROYUKI SHIMIZU
National Institute of Infectious Diseases
Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Chemical Engineering, Chemistry & Bioengineering Chemistry Immunology & microbiology Medicine & Pharmacology

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Motif Ii of Japanese Encephalitis Vius Ns3 Pro is Not Essential for Rna Binding Activit Utama, Andi; Shimizu, Hiroyuki
ANNALES BOGORIENSES Vol 11, No 1 (2007): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (2930.744 KB) | DOI: 10.1234/19

Abstract

The role of motif n  (DEAR AsPw-GlulK,,-Ala.,.7-Hi  1~~  )  of Japanese encephalitis  virus (JEV) NS3 protein on RNA binding activity was studied. A point mutation was introduced to  the motif and  the RNA binding activity of each mutant protein was analyzed. Truncated form of each protein with a His-tag was expressed m Escherichia coli BL2l(DE3) pLysS and purified by metal affinity resin. Asp-285 - and Glu-286 was respectively substituted with Ala, AJa-287 was replaced by Cys.,Gly.  or ser. His-288 was mutated to other 19 amino acids. In total. 24 mutant proteins were produced and analyzed. AL results, all mutants showed quite similar RNA binding activity.  Indicating that motif II of JEY NS3 is not related to RNA binding activity. The same finding was reported for hepatitis C virus c (Hey) NS3 protein, suggesting the similar structure of NS3 protein in flavivirus .Keywords: Japanese encephalitis, R.binding, motif
Construction Of A Recombinant Virus Between Poliovirus And Coxsackie A Virus 11 Utama, Andi; Shimizu, Hiroyuki
ANNALES BOGORIENSES Vol 10, No 1 (2005): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (3495.894 KB) | DOI: 10.1234/14

Abstract

Recent outbreaks of circulating vaccine-derived polio virus (cVDPV) revealed the possibility of recombination between vaccine strains poliovirus (PV) and cluster C enterovirus . Based on genetic analysis, it is assumed that coxackie virus 11 (CAV-11 ), one of the cluster enterovirus, may naturally recombine with PV. To elucidate this hypothesis , the chimeric virus between PJ156, a type 1 cVDPV isolate isolated from an acute flaccid paralys is (AFP) case in the Philippines in 2001 , and CAV-11 (PJI56/CAV-11) was constructed by using long-PCR method . As the result , PJl56/CAV-11 was aviable in HEp-2 cell line. The PJl 56/CAV-11 exhibited mostly similar phenotype with parental PJ156 in term of plaque size, viral growth and neurovirulence. These results suggested that recombination between PV and CAV-11 might naturally occur during transmission of vaccine strains in the community. The effect of recombination on the viral phenotype is significantly depending on the counterpart virus  
Co-Authors ANDI UTAMA