Development of genome-wide polymorphic microsatellite markers for Trichinella spiralis.

Background: Trichinella nematodes are globally distributed food-borne pathogens, in which Trichinella spiralis is the most common species in China. Microsatellites are a powerful tool in population genetics and phylogeographic analysis. However, only a few microsatellite markers were reported in T.

Newly excysted juveniles of Fasciola gigantica trigger the release of water buffalo neutrophil extracellular traps in vitro.

Polymorphonuclear neutrophils (PMNs) are the most abundant leukocytes and are among the first line of immune system defense. PMNs can form neutrophil extracellular traps (NETs) in response to some pathogens. The release of NETs plays an important role in trapping and killing invading parasites.

[Screening and Applications of qPCR Primers for apomucin Gene of Echinococcus multilocularis].

Objective: To screen for the optimal qPCR primers for Echinococcus multilocularis apomucin gene (Em-apo) and analyze Em-apo expression.

Methods: Primers were designed based on 4 Em-apo sequences from GeneDB. Primer specificity and PCR efficiency were determined, based on which the optimal primer pairs were selected.

[Identification, Expression and Antigenicity Analysis of Serpin B6 of Taenia solium].

Objective: To identify and express serpin B6 of Taenia solium (Tsserpin B6) and explore its possible use as a diagnostic antigen.

Methods: Primers for Tsserpin B6 were designed according to T. solium genome and transcriptome data.

[Identification of Taenia solium Insulin Receptor TsIR-1316 and Expression of Its Ligand Binding Domain].

Objective: To characterize the structure of insulin receptor of Taenia solium（TsIR-1316） and express its ligand binding domain (LBD).

Methods: Primers for TsIR-1316 were designed according to the genomic data of T. solium, and the TsIR-1316 gene was amplified by PCR.

Molecular identification of Diphyllobothrium latum and a brief review of diphyllobothriosis in China.

Two tapeworm specimens collected in northeast China in 2009 and 2011 were identified as Diphyllobothrium latum based on morphological criteria. Molecular methods were used to confirm their identity and analyze genetic variations compared with published data for this species. Species identity was confirmed by molecular characterization of the 18S rDNA partial sequence, complete sequences of internal transcribed spacers (ITSs) and 5.

Mapping codon usage of the translation initiation region in porcine reproductive and respiratory syndrome virus genome.

Background: Porcine reproductive and respitatory syndrome virus (PRRSV) is a recently emerged pathogen and severely affects swine populations worldwide. The replication of PRRSV is tightly controlled by viral gene expression and the codon usage of translation initiation region within each gene could potentially regulate the translation rate. Therefore, a better understanding of the codon usage pattern of the initiation translation region would shed light on the regulation of PRRSV gene expression.

[Construction, expression and immunogenicity of eukaryotic vectors based on goat pox virus P32 gene].

The full-length P32 gene and the truncated P32 gene (MP-32) were amplified from the recombinant plasmid pMD-P32 by polymerase chain reaction (PCR) and cloned into pcDNA3. 1(+) and pcDNA3.1-CpG respectively.

Rapid detection of porcine circovirus type 2 by loop-mediated isothermal amplification.

A method of loop-mediated isothermal amplification (LAMP) was employed to develop a rapid and simple detection system for porcine circovirus type 2 (PCV2). The amplification could be finished in 60 min under isothermal condition at 64 degrees C by employing a set of four primers targeting the cap gene of PCV2. The LAMP assay showed higher sensitivity than the conventional PCR, with a detection limit of five copies per tube of purified PCV2 genomic DNA.

[Molecular relationship of Eurytrema coelmaticum inferred from 18S rRNA sequence].

Objective: To elucidate the taxonomic position of Eurytrema coelmaticum by using molecular technology.

Methods: 18S rRNA fragment was amplified from E. coelmaticum genomic DNA by specific conservative primers and sequenced.

[Combined expression of TSO45W-4BX from Taenia solium and porcine CD58 in Escherichia coli].

Objective: To express the TSO45W-4BX of Taenia solium in combination with CD58 as a molecular adjuvant for improving the protective efficacy of the TSO45W-4BX recombinant vaccine.

Methods: TSO45W-4BX and porcine CD58 genes were amplified by PCR, using recombinant plasmids pGEM-4B and pGEM-CD58 as template respectively. The CD58 fragment was inserted into the recombinant plasmid pGEX-4T-1 with directly ligated TSO45W-4BX.

[Defense mechanisms of Taeniidae against host immune response].

The defense mechanisms of Taeniidae against host immune reaction were reviewed. The parasites may defend themselves from the host's immune attack by: (1)producing specific biochemicals as barriers against the damage caused by immune reactions, (2) changing surface antigens and secreting some active substances that interfere and deconstruct host's immune system and other hazards, (3) self-disguising through synthesizing homologies to host's substances in structure or function in order to avoid the immune surveillance of the host.

[Adjuvant effect of CpG DNA recombinant plasmid on antigen of Cysticercus cellulosae].

In order to prove the adjuvant effect of CpG DNA recombinant plasmid, the total antibodies and their IgG2a subtype induced by antigen of Cysticercus cellulosae, and content of IL-4 and IFN-gamma secreted from splenic cell of mouse immunized were measured. The recombinant plasmids showed an adjuvant effect, and CpG2 was the best adjuvant among the plasmids. It is proved that the CpG DNA possesses a synergistic effect with AI(OH)3 and 206 adjuvant, and is an effective Th1 type adjuvant in mice.

[Expression of Taenia solium oncosphere 45WB2 gene in Pichia pastoris system].

Objective: To amplify and express the gene from Taenia solium oncosphere in Pichia pastoris system.

Methods: Total RNA was extracted from hatched and activated oncospheres. A 459bp specific fragment was amplified by RT-PCR.

[High-level expression of the potential vaccine antigen TSO18 of Taenia solium in Pichia pastoris].

TSO18 gene was subcloned into the Pichia pastoris expression vector pPIC9K. The recombinant plasmid pPIC9K-TSO18 was transformed into P. pastoris GS115 by electroporation so that the plasmid will be integrated with chromosome of P.

[Screening of cDNA clone for putative RNA polymerase subunit of Cysticercus cellulosae].

Objective: To obtain related genes of Cysticercus cellulosae from spliced leader (SL) cDNA library.

Methods: Spliced leader library of Cysticercus cellulosae was constructed using SL specific primer and oligo (dT) 15 with M13M4 primer, and positive clones were then screened randomly, identified with enzyme restriction, followed by sequencing and homologous analysis.

Results: The amino acid sequence, encoded by the positive clone with a poly (A)22 tail and a complete open reading frame (ORF), was with homology of RNA polymerase subunit genes of human, B.