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

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

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

The human cytomegalovirus(HCMV) gene encoding the protein reactive with the sera of HCMV-infected patient was cloned and characterized. A reactive phage clone was screened from a lambda gt11 expression library of cDNA of HCMV AD169 strain using HCMV-infected patient sera. The recombinant protein was expressed as 138 kDa-fusion protein with beta-galactosidase, which was reactive with IgM or IgG HCMV antibody-positive sera, but not with anti-HCMV antibody-negative sera. A homology search of the DNA sequence of the cloned gene with HCMV AD169 sequences revealed that it was composed of 709 base pairs spanning between 0.174 and 0.177 map units of the UL32 region of the HCMV AD169 strain genome. This position corresponded to a part of the gene encoding 150 kDa phosphoprotein-(pp150), a major tegument protein, which was reported as an immunogenic protein which evoked strong and longstanding antibody response and had no sequence homology with the proteins of other herpesviruses. These results suggested that pp150 was an immunogenic protein in natural HCMV infection and therefore this clone was regarded as a useful candidate for developing an antigen for the serodiagnosis of HCMV.
Antibodies, Viral/*blood/immunology ; Antigens, Viral/*genetics/immunology ; Cloning, Molecular ; Cytomegalovirus/genetics/*immunology ; Cytomegalovirus Infections/blood/*immunology/virology ; DNA, Complementary/genetics ; DNA, Viral/genetics ; Gene Library ; *Genes, Structural, Viral ; Human ; Recombinant Fusion Proteins/biosynthesis/immunology ; Sequence Homology, Nucleic Acid ; Support, Non-U.S. Gov't ; Viral Matrix Proteins/*genetics/immunology

To develop a recombinant lentivirus co-expressing structural protein of hepatitis C virus (HCV) and secreted Gaussia Luciferase (Gluc), we first constructed an expression vector that encoded HCV structural protein (C, E1, E2) and GLuc named pCSGluc2aCE1E2. The expression of HCV proteins and Gluc was confirmed by an immunofluorescence assay (IFA) and the detection of luciferase activity. Recombinant lentivirus (VSVpp-HCV) was developed by the co-transfection of pCSGluc2aCE1E2 into 293T cells with pHR'CMVA8.2 and pVSVG. The infectivity of VSVpp-HCV was confirmed by luciferase activity detection, IFA and western blotting. Virus-like particles were identified using electron microscopy after concentration. The results showed that the level of luciferase activity correlated with the expression of HCV protein after the infection of cells with lentivirus VSVpp-HCV. Therefore, the expression level of HCV proteins could be evaluated by detecting the luciferase activity of Gluc. In conclusion, this research pave a way for the development of transgenic mice that express HCV proteins and Gluc, which enable the evaluation of anti-HCV therapy and vaccine in vivo.
Animals ; Copepoda ; Genes, Reporter ; Genetic Vectors ; genetics ; metabolism ; Hepacivirus ; genetics ; metabolism ; Hepatitis C ; virology ; Humans ; Lentivirus ; genetics ; metabolism ; Luciferases ; genetics ; metabolism ; Recombinant Fusion Proteins ; genetics ; metabolism ; Viral Structural Proteins ; genetics ; metabolism