Induction of Hantaan virus-specific immune responses in C57BL/6 mice by immunization with a modified recombinant adenovirus containing the chimeric gene, GcS0.7.

Hantavirus glycoprotein Gc is one of the main components that contribute to the generation of humoral immune responses, while the nucleocapsid protein (NP) is involved in cellular immune responses through the induction of antibody-dependent cytotoxic T cells. In this study, a chimeric gene, GcS0.7, which encodes a fusion protein containing Gc and truncated NP, was constructed as a candidate for Hantaan virus (HTNV) vaccine development.

Modification of the adenoviral transfer vector enhances expression of the Hantavirus fusion protein GnS0.7 and induces a strong immune response in C57BL/6 mice.

Hantavirus glycoproteins (Gn and Gc) are significant components of vaccines for haemorrhagic fever with renal syndrome (HFRS); however, they are not effective due to weak immunogenicity and low levels of production in expression systems. To circumvent this problem, a 0.7-kb fragment of the S segment was fused to Gn, and a hybrid CAG promoter/enhancer in conjunction with (or without) the WPRE (Woodchuck hepatitis virus post-transcriptional regulatory element) was used to improve the expression of fusion protein GnS0.

Soluble expression and purification of Brucella cell surface protein (BCSP31) of Brucella melitensis and preparation of anti-BCSP31 monoclonal antibodies.

Brucella cell surface protein (BCSP31) is potentially useful for diagnosing brucellosis. We aimed to establish a monoclonal antibody (MAb) against Brucella melitensis BCSP31 and to investigate its distribution in diagnosis. Soluble recombinant BCSP31 was successfully expressed and purified.

[Construction and immunogenic study of recombinant adenovirus containing chimeric gene G2S0.7 and CTL epitopes of Hantaan virus].

Aim: To express G2 fragment of M segment and 0.7 kb fragment of S segment and several CTL epitopes of S segment in adenovirus expression system and investigate the immunological properties of hantaan virus chimeric gene.

Methods: The recombinant adenovirus was constructed and the recombinant adenovirus was obtained after transfecting HEK293 cells.

Downregulation of NDRG1 promotes invasion of human gastric cancer AGS cells through MMP-2.

The N-myc downstream-regulated gene-1 (NDRG1) has recently been proposed as a metastasis suppressor, but its precise role remains unclear. To investigate whether NDRG1 can indeed influence the metastasis progress, expression of endogenous NDRG1 was knocked down in human AGS gastric adenocarcinoma cells using RNA interference. Stable NDRG1 "silenced" transfectants showed similar growth rates as their control counterparts.

[Construction and identification of a new type of recombinant adenovirus containing S0.7 gene of Hantavirus].

Aim: To construct a adenovirus vector containing the 0.7 kb fragment of S gene of Hantavirus, CAG promoter, and WPRE (mRNA-stabilizing post-transcriptional regulatory element from the woodchuck hepatitis virus).

Methods: The fragments of CAG and WPRE were synthesized according to GenBank, and inserted into the plasmid pShuttle-S0.

[Transformation of full-length gene of mouse/human chimeric antibody against Hantaan virus into Arabidopsis thaliana].

Aim: To construct the transgenic Arabidopsis thaliana containing full-length gene of mouse/human chimeric antibody(3G1MH) against Hantaan virus.

Methods: The recombinant plasmid 3G1MH-pCAMBIA2301 was transformed into Agrobacterium tumefaciens GV3101 by TSS freeze-thaw method, and then the recombinant was transferred into wild Arabidopsis thaliana by vacuum-transgenic method. The regenerated transgenic plants were selected with kanamycin, and confirmed by PCR and Northern blot.

Short hairpin RNA targeting survivin inhibits growth and angiogenesis of glioma U251 cells.

Survivin is a novel tumor-associated gene, its overexpression mostly associates with carcinogenesis and development. Nevertheless, the precise role of survivin in initiation and progression of gliomas is still not completely clear. We constructed here three short hairpin RNA (shRNA) targeting survivin plasmid vectors and introduced them into glioma U251 cells.

The expression and genetic immunization of chimeric fragment of Hantaan virus M and S segments.

Hemorrhagic fever with renal syndrome (HFRS), which is characterized by severe symptoms and high mortality, is caused by hantavirus. There are still no effective prophylactic vaccines directed to HFRS until now. In this research, we fused expressed G2 fragment of M segment and 0.

[The effect of HCV core protein on the expression of cyclooxygenase 2 (COX-2) in HepG2 cells].

Aim: To investigate the effect of Hepatitis C virus (HCV) core protein on the expression of cyclooxygenase 2 (COX-2).

Methods: The genes encoded HCV core protein were amplified from plasmid containing full length genome of HCV strain H77 using PCR, and were cloned into eukaryotic expression vector pcDNA3.1.

[Expression and genetic immunization of hantaan virus G2 recombinant adenovirus].

Aim: To express hantaan virus(HTNV) envelope glycoprotein G(2) recombinant adenovirus(Adeno-G(2)) in vero E6 cells and explore its property of inducing immune response.

Methods: Vero E6 cells were infected with the HTNV Adeno-G(2) (100 MOI). The expression of Adeno-G(2) in the infected Vero E6 cells was detected by IFA.

[Transient expression and characterization of intracellular single chain Fv against the nucleocapsid protein of Hantavirus].

Aim: To transiently express an intracellular single chain Fv of monoclonal antibody 1A8 against nucleocapsid protein of Hantavirus and characterize the immunological activities of the expressed products.

Methods: COS-7 cells were transfected with mammalian expression vector 1A8-scFv-Ckappa/pCI-neo via lipofectin. The expressed product was identified by indirect immunofluorescence and immunoprecipitation.

[Expression of amino terminal of nucleoprotein and glycoprotein G2 of Hantaan virus in insect cells in the form of fusion protein].

Aim: To express the glycoprotein G2 and amino terminal of nucleoprotein (NP) of Hantaan virus in Bac-to-Bac baculovirus expression system in the form of fusion protein.

Methods: The recombinant baculovirus expression vector pFBDHTa-G2S 0.7 was constructed.