The objective of the present study was to express recombinant SjPDIA3 protein in E.coli and prepare polyclonal antibodies. The PDIA3 gene was amplified in PCR using the genome of Schistosoma japonicum as a template. The PCR products obtained were inserted into prokaryotic expression vector pET28a(+). The recombinant plasmid pET28a(+)-SjPDIA3 was transformed into E.coli BL21(DE3). Subsequently, SjPDIA3 protein was expressed with induction of IPTG and purified using NI-NTA resin. The BALB/c mice were immunized with recombinant SjPDIA3 protein to generate polyclonal antibodies. The results showed that a cDNA of 1482 bp was amplified. Bioinformatics analysis determined that the full-length ORF was 1482 bp encoding 493 amino acids and SjPDIA3 protein had a thiol-disulfide oxidoreductase domain and a signal peptide with no transmembrane peptide. Western blot demonstrated prokaryotic expression of the recombinant SjPDIA3 protein with molecular mass of 55 kDa and its efficient antigenicity and immunogenicity as expected.

The present study aimed at obtaining the recombinant SjCHGC06822 protein by using prokaryotic expression system and evaluating the immune response in BALB/c mice. A full-length ORF encoding SjCHGC06822 of S.japonicum was amplified in PCR. The obtained cDNA was subcloned into pET28a(+) vector that was then transformed into BL21(2E). The recombinant protein was purified using His-Ni resin. Bioinformatics analysis showed that the full-length ORF was 591 bp, encoding 192 amino acids. The SjCHGC06822 protein had an EF-hand domain without signal peptide and transmembrane peptide. The recombinant protein was visualized as a band at molecular mass of 23 kDa in Western blot. BALB/c mice were immunized with the recombinant protein and antibody responses were detected in ELISA. The immunized mice along with non-immunized controls were challenged with S.japonicum and worm reduction rate and liver egg reduction were recorded. In two separate animal experiments, worm reduction rates of the immunized mice were 30.36%and 40.26%and liver egg reductions were 30.14%and 12.28%. The specific IgG, IgG2a and IgG1 were detected using ELISA. The antibody responses gradually increased post immunization and subsequent challenge. However, IgG2a and IgG1 levels increased post immunization but decreased post challenge while the ratio of IgG2a/IgG1 (<1) was climbing. In conclusion, the recombinant SjCHGC06822 protein can induce good immunogenicity.

为研究Wnt信号通路对血吸虫发育的调节作用,本研究克隆鉴定了日本血吸虫的Wnt1基因(SjWnt1).序列分析显示,SjWnt1蛋白具有Wnt信号蛋白家族的结构功能域,但比高等生物的Wnt1少一个保守的Cys.mRNA和蛋白的定量分析结果显示,SjWnt1表达于血吸虫的整个发育过程,而在虫卵和早期童虫中呈高表达水平,提示SjWnt1对卵胚发育和早期童虫的细胞分化具有调节作用.来源于非适宜宿主及单性感染的发育不良虫体中SjWnt1 mRNA水平比发育正常的虫体高,表明不良发育的虫体相应的Wnt信号阻滞.SjWnt1表达下调后则引起Wnt/β-catenin、Wnt/PCP及Wnt/Ca2+通路的相应下游基因的mRNA水平下降,推测SjWnt1可能通过这3种传递途径行使信号传递的功能.

The objective of the present study was to express recombinant SjHSP70 in insect cells Sf9. The SjHSP70 gene of 1962 bp was amplified by PCR and used to construct vector pFastBacHTA-SjHSP70. Then, reBacmid-SjHSP70 was transformed into competent cells. The recombinant shuttle plasmid reBacmid-SjHSP70 was obtained from the transformed DH10Bac cells and then transfected into insect cells Sf9. The recombinant protein SjHSP70 expressed in infected cells was visualized in IFA and purified with His column affinity chromatography. Western blot demonstrated the ecombinant SjHSP70 was recognized by antiserum from mice infected with Schistosoma japonicum. The availability of recombinant SjHSP70 provided materials for further exploring its immunobiological function.