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目的 探讨大鼠抗日本血吸虫 (Schistosomajaponicum ,Sj)感染的分子机制 ,为血吸虫病疫苗的研制提供新的抗原分子。方法 用Sj感染大鼠血清筛选Sj成虫cDNA文库 ,对阳性克隆cDNA插入片段进行PCR扩增及测序分析。 结果 对cDNA文库中约 3× 10 5个噬菌斑进行筛选 ,获 7个阳性克隆 ,其插入基因片段大小为 0 9kb～ 2 0kb。初步测序获 5个部分序列 ,其中R3与Sj线粒体基因明显同源 ;R5为新基因序列 ,命名为Sj-Cs2 (GenBank的登录号为AY0 36 5 80 ) ,经计算机分析该基因片段编码 2 0 6个氨基酸组成的跨膜蛋白 ,Sj-Cs2蛋白含有 3个蛋白激酶C磷酸化位点和 6个N -肉豆蔻酸化位点 ;其余序列亦为新基因片段 ,但无完整编码框。结论 筛选获得了与大鼠抗Sj感染有关的基因克隆 ,相关克隆cDNA全长的获取、新基因的结构与功能以及免疫学特性值得深入研究
Objective To investigate the molecular mechanism of rat anti-Schistosoma japonicum (Sj) infection and to provide a novel antigen molecule for the development of schistosomiasis vaccine. Methods Sj adult rat cDNA library was screened by Sj infected rat serum. The positive cloned cDNA inserts were amplified by PCR and sequenced. Results About 3 × 10 5 plaques were screened in the cDNA library. Seven positive clones were obtained, and the insert size ranged from 0 9 kb to 20 kb. Preliminary sequencing of the five partial sequences was made, of which R3 and Sj mitochondrial genes were significantly homologous; R5 is a new gene sequence, named Sj-Cs2 (GenBank accession number AY036 5 80), the computer analysis of the gene fragment encoding 20 6 amino acid transmembrane protein, Sj-Cs2 protein contains 3 protein kinase C phosphorylation sites and 6 N-myristoylation sites; the rest of the sequence is also a new gene fragment, but no complete coding frame. Conclusion The cloning of the gene related to anti-Sj infection in rats was obtained by screening, and the full-length cDNA cloning, the structure and function of the new gene, and the immunological properties deserved further study