华支睾吸虫CsTMP63o基因的克隆、表达及感染者抗体亚类分析

Cloning,expression and specifi antibody analysis in infectants of the Cs TMP 63o of Clonorchis sinensis

目的:对华支睾吸虫跨膜蛋白63c基因进行克隆、表达,并通过感染者抗体亚类分析评价其免疫学诊断效果。方法:根据Cs TMP 63c(Gen Bank序列号:AF093242)基因序列,拼接其多个跨膜区片段,人工合成新基因Cs TMP 63o,制备抗原免疫小鼠,检测抗体效价,建立抗体检测系统,初步分析感染者针对Cs TMP 63o的特异性抗体亚类。结果 :从Cs c DNA文库中筛选出Cs TMP 63c基因,全长2 013 bp,由大肠杆菌表达、纯化,得到分子量为38 k D的重组蛋白。免疫大鼠后,抗体效价较低;感染者的抗体亚类分布以IgG4类为主,与各项常规生化和血液指标未发现有显著相关性。结论 :Cs TMP 63o在大肠杆菌中高效表达,具有较好的免疫原性,但血清诊断价值不理想。

Objective To clone and express the CsTMP 63o gene, in order to analyze the immunological diagnosis effect on the infecters＇ antibody subclasses. Methods According to CsTMP 63c （GenBank accession： AF093242） gene sequences, multiple transmembrane region were sliced into fragments to synthesize the new gene CsTMP 630. Mice with antibody were prepared, and the antibody were evalusted. Antibody mechanism was established to analyze CsTMP 630 specific antibody subgroup. Results A full-length 2013 bp cDNA clone of CsTMP 63c was selected, and expressed and purified by Escherichia coll. Recombinant protein weight was 38kD. After mice were immunized, the antibody titer was low. The antibody subtype was mainly IgG4 and has no significant correlation with all biomedical and blood indexes. Conclusion CsTMP 63o is highly expressed in E. coli, with better immunogen, but the diagnostic value of serum is not ideal.