[Subcloning of M1 gene fragment of H5N1 influenza virus and its expression in Escherichia coli].

Objective: To generate the Escherichia col vector expressing human H5N1 influenza virus M1 protein. To provide useful tools for detection of human H5N1 influenza virus and study on biological function of M1 protein.

Methods: M1 gene fragment was amplified by PCR using the influenza virus gene segment 7 as template, and was subcloned into pQE80-L vector.

[Construction and immunogenicity study of DNA vaccine expressing human H5N1 influenza virus hemagglutinin].

Based on the first isolated human H5N1 influenza virus strain A/Anhui/1/2005 in China, HA and HA1 genes were amplified and cloned into the eukaryotic expression vector pStar. The recombinant plasmids pStar-HA and pStar-HA1 were transfected into COS7 cells. Western blot and IFA showed that the two recombinant DNA plasmids were successfully expressed in eukaryotic cells.

[Genetic analysis of NS1 fragment of human H5N1 influenza virus isolated in Anhui province and its expression in Escherichia coli].

NS1 gene was amplified from an H5N1 influenza virus, A/Anhui/01/2005 for cloning, sequence analysis and later expression in Escherichia coli. The result showed that NS1 of A/Anhui/01/2005 strain had the close phylogenetic relationship with that of H5N1 avian influenza strains recently isolated in Fujian and Hunan. Correspondingly, its amino acid sequence showed the highest homology with that of Fujian and Hunan strains.

[Expression of sTNFR-IgGFc fusion gene in endothelial cell and its application in gene therapy for rheumatoid arthritis].

Tumor necrosis factor alpha (TNFalpha) is a pro-inflammatory cytokine, acting as a regulator of inflammation and immunity. TNFalpha plays a critical role in the pathogenesis of rheumatoid arthritis. Blocking of TNFa activity suppressed inflammatory tissue damage.

[Screening for new binding proteins which interact with BM2 of influenza B virus with yeast two-hybrid system].

Objective: To explore the role of BM2 protein in the life cycle of influenza B virus.

Methods: The authors screened human kidney MATCHMAKER cDNA library for new binding partners of BM2 of influenza B virus by using the yeast two hybrid system with truncated BM2 (26-109 aa) as the bait.

Results: Six positive plasmids encoding N-acetylneuraminate pyruvate lyase, angiopoietin 3, zinc finger protein 251, ribosomal protein S20, protein arginine N-methyltransferase 1 variant 1 (PRMT) and transcription factor-like 1 (TCFL1) were obtained.

[Evaluation on the biosafety of classical swine fever DNA vaccine].

The biosafety of DNA vaccine is one of the key questions which should be solved before it is used in the clinical trail. In order to evaluate the biosafety of DNA vaccine, the CSFV DNA vaccine was used in the studying target, two main aspects of the vaccine were explored in the study. Firstly, the possibility of integration of two kinds of DNA vaccine plasmids into pig genome was analyzed by PCR technology after the different vaccines were injected through the intramuscular introduction.