[Co-expressed GP5 and M proteins of porcine reproductive and respiratory syndrome virus can form heterodimers].

In order to investigate the characterization of in vitro co-expressed GP5 and M proteins of porcine reproductive and respiratory syndrome virus (PRRSV), eukaryotic expression plasmids pCI-ORF5 (expressing GP5 protein alone), pCI-ORF6 (expressing M protein alone), and pCI-ORF5/ORF6 (co-expressing GP5 and M proteins) were constructed. After transient transfection, Western blot analysis under nonreducing condition demonstrated that co-expressed GP5 and M proteins could form disulfide-linked heterodimers (GP5-M) in transiently transfected BHK-21 cells. To further study the influence of GP5-M heterodimers formation on the subcellular localizations of GP5 or M proteins, green fluorescence protein (EGFP) and red fluorescence protein (RFP) were used as markers. The results of fluorescence distribution showed that co-expressed GP5-EGFP chimera and M-RFP chimera boosted the transport of GP5 from the endoplasmic reticulum (ER) to the Golgi complex, indicating that the formation of GP5-M heterodimers may be involved in posttranslational modification, transport, and subcellular localization of GP5. These results presented here lay foundation to further study the molecular mechanism of GP5-M heterodimer formation and its role in protective immunity of PRRSV.

Mu1,1-azide-bridged ferromagnetic MnIII dimer with slow relaxation of magnetization.

A new end-on azide-bridged dimeric Mn(III) complex has been synthesized by using the tridentate N-isonicotinamidosalicylaidimine ligand. Magnetic studies show that the complex has a high-spin ground state of S = 4 and is ac out-of-phase frequency-dependent.

mer-[Fe(pcq)(CN)3]-: a novel cyanide-containing building block and its application to assembling cyanide-bridged trinuclear FeIII2MnII complexes [pcq- = 8-(pyridine-2-carboxamido)quinoline anion].

A new cyanide-containing building block K[Fe(pcq)(CN)(3)] [1; pcq(-) = 8-(pyridine-2-carboxamido)quinoline anion] containing a low-spin Fe(III) center with three cyanide groups in a meridional arrangement has been successfully designed and synthesized. Three cyanide-bridged trinuclear Fe(III)(2)Mn(II) complexes, [Fe(pcq)(CN)(3)](2)[Mn(CH(3)OH)(2)(H(2)O)(2)].2H(2)O (2), [Fe(pcq)(CN)(3)](2)[Mn(bipy)(2)].CH(3)OH.2H(2)O (3), and [Fe(pcq)(CN)(3)](2)[Mn(phen)(2)].CH(3)OH.2H(2)O (4), have been synthesized and structurally characterized. The magnetic susceptibilities of the three heterometallic complexes have been investigated.

[Immunogenicity of DNA vaccine expressing GP5 of porcine reproductive and respiratory syndrome virus fused with VP22 of bovine herpesvirus 1].

To enhance the immuogenicity of DNA vaccines expressing the GP5 protein of Porcine Reproductive and Respiratory Syndrome Virus (PRRSV), the tegument protein VP22 (encoded by VP22 gene) of Bovine Herpesvirus 1 (BHV-1), which has been demonstrated to exhibit the unusual protein transduction property, was fused to N-terminus of GP5 of DNA vaccine construct pCI-ORF5M, resulting in pCI-VP22-ORF5M expressing VP22-GP5 fusion protein. The expression of VP22-GP5 fusion protein was confirmed by both indirect immunofluorescence assay (IFA) and Western blot. To investigate its immunogenicity, BALB/c mice were immunized with the fusion expression plasmid pCI-VP22-ORF5M and non-fusion expression plasmid pCI-ORF5M, respectively. The GP5-specific ELISA antibodies, neutralizing antibodies and lymphocyte proliferative responses were evaluated at various time points after primary immunization. The results showed that GP5-specific ELISA antibodies, neutralizing antibodies, and lymphocyte proliferative responses induced by DNA vaccine pCI-VP22-ORF5M were higher significantly than those of DNA vaccine pCI-ORF5M, indicating that fusion expression with BHV-1 VP22 significantly enhances the immuogenicity of DNA vaccine expressing the PRRSV GP5 protein, and that this strategy may also be useful to develop more efficient DNA vaccines against other pathogens.

[Expression of GP5-M fusion protein of porcine reproductive and respiratory syndrone virus (PRRSV) and establishment of ELISA diagnose based on the recombinant fusion protein].

The cDNA fragment encoding the truncated GP5 and the full-length M protein of Porcine Reproductive and Respiratory Syndrone Virus (PRRSV) were orderly fused to the downstream of glutathione S-transferase (GST) of pGEX-KG expression vector, resulting in the fusion expression plasmid pKG-56. After transformed into E. coli BL21 (DE3) and induced by IPTG, the results of SDS-PAGE showed that the GST-GP5-M fusion protein was expressed in high level. Western-blot was performed to confirm that the expressed fusion protein could specifically react with antiserum against PRRSV. The fusion protein was further purified and used as an antigen to establish a novel PRRSV ELISA diagnose assay (P56-ELISA). Comparison between P56-ELISA and the abroad kit IDEXX-ELISA showed the two methods had 94.1 percent agreement by detecting 205 serum samples, indicating that the indirect P56-ELISA was specific and sensitive. The correlation between virus neutralization antibody of the infected pigs (not convalescent pigs) and antibody response to the fusion protein GP5-M was further studied. The regression function analysis suggested that there was no significant correlation between ELISA antibody response (OD630 nm) to the fusion protein GP5-M in clinical serum and their specific neutralizing titers.

Synthesis, crystal structure, and magnetic properties of oxime-bridged polynuclear Ni(II) and Cu(II) complexes.

Two new polynuclear complexes [Ni6(amox)6(mu6-O)(mu3-OH)2](Cl2).6H2O and [Cu3(amox)3(mu3-OH)(mu3-Cl)](ClO4).4H2O (amox- = anion of 4-amino-4-methyl-2-pentanone oxime) have been synthesized and characterized structurally and magnetically. The Ni(II) complex contains a novel Chinese-lantern-like Ni6 cage centered by an oxo ion. It contains the nearest octahedral Ni(II)...Ni(II) separation (<2.8 A) and exhibits strong antiferromagnetic properties. The Cu(II) complex has a cyclic trinuclear copper(II) core bridged by both mu3-OH(-) and mu3-Cl(-) ions. The magnetic susceptibilities of both antiferromagnetic complexes were fitted by using approximate models.

[Construction and immunogenicity of recombinant pseudorabies virus expressing the modified GP5m protein of porcine reproduction and respiratory syndrome virus].

Pseudorabies virus (PRV), an alpha-herpesvirus, has been used as a vector for live-viral animal vaccines. The recombinant PRV TK- / gE- / GP5+, which expressing GP5 of PRRSV, is developed based on the PRV genetic-depleting vaccine-virus strain, TK- / gE- /LacZ+. However, this strain stimulated poorly the vaccinated animals to produce neutralizing antibodies against PRRSV. In order to develop a booster specific immunized response of the PRV recombinant, the ORF5 gene of PRRSV TK- / gE- / LacZ+ was substituted by a modified ORF5 gene, ORF5m. The resultant recombinant PRV, TK- /gE- / GP5m+, was verified by PCR, Southern blotting and Western blotting. TK- / gE- / GP5m+ and TK- / gE- / GP5+ expressed GP5 proteins were inoculated into balb/c mice to evaluate their immunogenicity. The results demonstrated that the amount of neutralization antibodies and cell-immunity responses induced by TK- / gE- /GP5m+ against PRRSV were higher than that of TK- / gE- / GP5+. This study indicated that the new recombinant PRV expressing the modified GP5m protein is a candidate for the development of bivalent genetic engineering vaccines against PRRSV and PRV.

A novel thiocyanate-bridged one-dimensional chain complex: [Cu(NCS)2(Hambi)] (Hambi is 2-aminomethyl-1H-benzimidazole).

Using 2-aminomethyl-1H-benzimidazole as the ligand, a new thiocyanate-bridged copper(II) complex, namely bis(2-aminomethyl-1H-benzimidazole-kappa(2)N(2),N(3))dithiocyanatocopper(II),[Cu(NCS)(2)(C(8)H(9)N(3))], has been synthesized and structurally characterized. The Cu atom is five-coordinated and exhibits a distorted square-pyramidal geometry. The thiocyanate ions (NCS(-)) act as either bridging or terminal ligands. The bridging NCS(-) ligands connect neighboring Cu atoms, constructing chains, while the terminal NCS(-) ligands form hydrogen bonds with amine H atoms, leading to a complicated network.

Cyano-bridged 2D Cu(II)-Cr(III) coordination polymers: structural evidence for formation of a polymeric macrocyclic metallic compound.

Two unique cyano-bridged 2D coordination polymers have been synthesized and characterized structurally and magnetically. The complexes contain two polyaza Cu(II) units and one novel macromolecular Cu(II) moiety, which have been synthesized via one-pot metal template condensation reactions involving ethylenediamine (en) and formaldehyde. Self-assembly of the polyaza Cu(II) mixture with [Cr(CN)(6)](3)(-) gave rise to two layered complexes. One complex contains unprecedented covalently linked polymeric Cu(II) chains and cyano-bridged Cu(II)(-)Cr(III) coordination chains, which are interwoven to form a novel layer. The other complex shows intriguing encapsulation of [Cr(CN)(6)](3)(-) anions. Intermetallic ferromagnetic coupling is operative within the bridged 2D layer. The magnetic susceptibilities of both complexes were simulated using approximate models.