Codon ; Dengue Virus ; chemistry ; genetics ; Gene Expression ; Genes, Viral ; Hydrophobic and Hydrophilic Interactions ; Viral Structural Proteins ; genetics

Animals ; Arteritis Virus, Equine ; chemistry ; genetics ; Horses ; Open Reading Frames ; Viral Nonstructural Proteins ; chemistry ; physiology ; Viral Structural Proteins ; chemistry ; physiology

OBJECTIVESTo establish an animal model of HCV transgenic mice to elucidate the pathogenesis of hepatitis C virus infection and function of the viral structural proteins.

METHODSStructural gene of HCV were amplified and recombined into eukaryotic expression vectors, pcDNA4HisMax and pMT/BiP/V5-His A, after their expressive activity was confirmed to detect the structural protein in the transfected COS7 and S2 cells by Western blot. The fertilized expression element, which contained CMV or pMT promoter, structural gene of HCV and polyadenylation signal sequence, was microinjected into 1736 C57BL/6 mouse fertilized ova. The ova were then replanted into the oviducts of 69 pseudopregnant recipient mice.

RESULTSTwenty-five recipient mice were impregnated and later produced 105 newborns; 49 of them died from unknown causes and 57 survived. After the specific HCV structural genes were identified by PCR and Southern blot hybridization, 26 founders were obtained; among them 10 were stable expression mice and 16 were the inducible ones. The rate of founders developed from implanted embryos was only 1.50%. Through hybridization with normal mice, 58 hybrid mice have been obtained at present.

CONCLUSIONTwo kinds of different transgenic mice of HCV were developed; one is of stable expression, and the other is inducible. This transgenic mice model may create an opportunity for studying the function of the structural gene of HCV and elucidate its pathogenicity.

Animals ; Disease Models, Animal ; Gene Expression Regulation, Viral ; Hepacivirus ; genetics ; Hepatitis C ; Mice ; Mice, Transgenic ; Viral Structural Proteins ; genetics

OBJECTIVETo investigate the know gene types of main wild type measles virus strains and take measures to control measles in Jilin Province.

METHODSGenetic characterization of 9 measles viruses isolated from 72 throat swabs or urine specimens of measles patients using CDW(150) cells line was studied in Jilin Province in 2005.

RESULTSSequence analysis of 450 nucleotides of COOH-terminal of nucleoprotein (N) genes of 9 isolates indicated that all were members of H(1) genotype, in which there are 7 strains of H1a and 2 strains of H1b, the H1a subgroup differed from H1b by 2.0% approximately 3.5% at the nucleotide level in the COOH-terminal of the N gene.

CONCLUSIONSThe H(1) genotype of wild-type measles viruses should be the main epidemic strain and main pathogen that caused measles outbreaks and sporadic cases in Jilin Province.

China ; Genes, Reporter ; Genes, Viral ; Genotype ; Humans ; Measles ; epidemiology ; virology ; Measles virus ; genetics ; Molecular Sequence Data ; Viral Structural Proteins ; genetics

Animals ; Genome, Viral ; Porcine respiratory and reproductive syndrome virus ; genetics ; pathogenicity ; Swine ; Viral Nonstructural Proteins ; genetics ; physiology ; Viral Structural Proteins ; genetics ; physiology ; Virulence ; genetics

In order to make clear the packing mechanism of the BmNPV polyhedra, a polyhedrin gene negative recombinant baculovirus, vBmBac(polh-)-5B-EGFP, expressing EGFP was constructed, and used to infect BmN cells jointly with wild-type BmNPV. Fluorescent microscopic observation demonstrated that EGFP and polyhedrin were expressed simultaneously, and the EGFP expression and polyhedra formation occurred in most of the jointly infected cells. Analysis of the purified polyhedra from jointly infected BmN cells showed that the foreign proteins were present in the polyhedra. The results indicated that BmNPV polyhedrin could incorporate proteins other than viral proteins into the polyhedra. It implies that a nonspecific recognition mechanism exists in the embedment of BmNPV polyhedra.
Animals ; Bombyx ; Gene Expression ; Green Fluorescent Proteins ; genetics ; metabolism ; Nucleopolyhedrovirus ; genetics ; physiology ; Viral Structural Proteins ; genetics ; metabolism ; Virus Assembly

OBJECTIVEGenetic evolution of VP1 of enterovirus type 71 in Shenzhen were analyzed.

METHODSAll samples were tested by RT-PCR using EV71 specific primer. The VP1 of EV71 were amplified and sequenced. A phylogenetic tree was constructed by comparison of the sequences with subgenotype A, B and C using DNAStar, BioEdit and Mega 3.1 software.

RESULTSAmong 35 strains, the homogeneity of the VP1 nucleotide sequence was between 92.1%-100%. The homogeneity of the VP1 nucleotide sequence with subgenotype A and B was between 81.4% -91.1%. The VP1 nucleotide sequence of 35 strains of Shenzhen shared between 93% -97.4% identity with cluster C4. The prevalence strains of EV71 were cluster C4b from 1998 to 2004, and gradually moved to C4a since 2003. All of EV71 were C4b from 2006 to 2008. Also, the homogeneity of the VP1 nucleotide sequence with Anhui FY23 EV71 strain were 94.5% -94.7%, 95.7% -95.8%, 96.2%, 95.4% -97.5%, 96.3% -99.2% from 2003 to 2008. It shows that the homogeneity was increased year by year. There was a mutation (A --> C) at No. 66 nucleotide of VP1 of EV71 that two strains were isolated in 2003 and 8 strains in 2008, that caused amino acid mutation (Q --> H) at No. 22 of VP1.

CONCLUSIONEV71 C4b was gradually moved to C4a from 1998 to 2008. There was a missense mutation at No. 66 nucleotide of VP1.

Enterovirus ; classification ; genetics ; Humans ; Mutation, Missense ; genetics ; Phylogeny ; Reverse Transcriptase Polymerase Chain Reaction ; Viral Structural Proteins ; classification ; genetics

The newly emerged Middle East respiratory syndrome coronavirus (MERS-CoV) is a highly pathogenic respiratory virus with pathogenic mechanisms that may be driven by innate immune pathways. The goal of this study is to characterize the expression of the structural (S, E, M, N) and accessory (ORF 3, ORF 4a, ORF 4b, ORF 5) proteins of MERS-CoV and to determine whether any of these proteins acts as an interferon antagonist. Individual structural and accessory protein-coding plasmids with an N-terminal HA tag were constructed and transiently transfected into cells, and their native expression and subcellular localization were assessed using Wes tern blotting and indirect immunofluorescence. While ORF 4b demonstrated majorly nuclear localization, all of the other proteins demonstrated cytoplasmic localization. In addition, for the first time, our experiments revealed that the M, ORF 4a, ORF 4b, and ORF 5 proteins are potent interferon antagonists. Further examination revealed that the ORF 4a protein of MERS-CoV has the most potential to counteract the antiviral effects of IFN via the inhibition of both the interferon production (IFN-β promoter activity, IRF-3/7 and NF-κB activation) and ISRE promoter element signaling pathways. Together, our results provide new insights into the function and pathogenic role of the structural and accessory proteins of MERS-CoV.
Cell Line ; Coronavirus ; genetics ; pathogenicity ; Genes, Viral ; Humans ; Interferons ; antagonists & inhibitors ; Open Reading Frames ; Recombinant Proteins ; genetics ; metabolism ; Viral Matrix Proteins ; genetics ; metabolism ; Viral Regulatory and Accessory Proteins ; genetics ; metabolism ; Viral Structural Proteins ; genetics ; metabolism

Rubella virus (RV), one of pathogens in TORCH syndrome, can lead to anisotropy of the fetus, and therefore it is of great significance to screen RV infection among women at child-bearing age. At the present, the screening of RV infection is mainly based on the ELISA method, the specificity of RV antigens is very important for ELISA tests. The antigenicity of RV is closely associated with its structural proteins, including capsid protein C, and envelope glycoproteins E1 and E2, which are important surface antigens of RV. This paper reviews the advances in the studies on the structure features, immunogenicity and recombinant proteins of the three structural proteins.
Antibodies, Viral ; analysis ; Enzyme-Linked Immunosorbent Assay ; Recombinant Proteins ; immunology ; Rubella virus ; genetics ; immunology ; Viral Core Proteins ; genetics ; immunology ; Viral Envelope Proteins ; genetics ; immunology ; Viral Structural Proteins ; genetics ; immunology

OBJECTIVETo obtain recombinant baculovirus containing full-length structural gene of HEV and to express HEV structural protein.

METHODSFull-length structural gene of HEV (5147-7126 nt) was inserted into baculovirus expression vector pAcUW51. The recombinant plasmid and Baculo Gold DNA were co-transfected into insect cell line Sf9. Single plaque was picked and amplified and expression of HEV structural protein was tested by SDS-PAGE, Western blot and immunofluorescence methods.

RESULTSSDS-PAGE analysis showed HEV structural protein was highly expressed; Western blot and immunofluorescence assay showed that the expression product could specifically react with HEV positive sera, confirming the protein possessing HEV specific antigenicity.

CONCLUSIONSHEV structural protein was successfully expressed using baculovirus expression system.

Animals ; Baculoviridae ; genetics ; Cells, Cultured ; Gene Expression ; Genes ; genetics ; Genetic Vectors ; Hepatitis E virus ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Viral Structural Proteins ; biosynthesis ; genetics