论文部分内容阅读
目的 筛选日本血吸虫 (Schistosomajaponicum ,Sj)新的疫苗候选基因或雌虫性别特异性基因。 方法 用Sj雌虫抗原免疫家兔制备血清 ,对Sj成虫cDNA文库进行免疫筛选 ,将阳性克隆的插入片段进行PCR扩增 ,采用Sanger双脱氧核苷酸末端终止法对阳性克隆插入子进行测序 ,与GeneBank中的已知序列进行比对分析。 结果 共获得 2 6个阳性克隆 ,其PCR产物大小约 0 .5～ 3kb。在进行测序的 8个阳性克隆中发现了GeneBank未报道的Sj肌球蛋白的部分基因序列 (GeneBank登录号为AY3 2 2 14 8)和 1个新基因 (命名为Sj F1,GeneBank登录号为AY2 61995 )。Sj F1编码的蛋白理论分子质量为 13 .45ku ,等电点为 6.2 9,富含α螺旋和随机卷曲 ,含多个蛋白激酶磷酸化位点 ,1个酪氨酸激酶Ⅱ磷酸化位点和 1个豆蔻酸位点。 结论 Sj雌虫免疫血清可识别Sj特异性抗原分子 ,该抗原基因是否为雌虫性别特异性基因及其作为日本血吸虫病疫苗候选基因的价值 ,有待进一步研究。
Objective To screen for new vaccine candidate genes or female sex-specific genes of Schistosoma japonicum (Sj). Methods Rabbit was immunized with Sj female antigens to prepare serum. The Sj adult cDNA library was screened by immunoprecipitation. The inserted fragment of positive clone was amplified by PCR. Positive cloned insert was sequenced by Sanger dideoxynucleotide termination method. Alignment with known sequences in GeneBank. Results A total of 26 positive clones were obtained. The size of PCR products was about 0.5-5 kb. A partial gene sequence of Sj myosin that was not reported by GeneBank (GeneBank accession number: AY222814) and a new gene (named as SjF1, GeneBank accession number: AY2 61995). Sj F1 encoded protein molecular weight of 13.45ku, isoelectric point of 6.29, rich in α-helix and random coil containing multiple protein kinase phosphorylation sites, a tyrosine kinase Ⅱ phosphorylation site and 1 myristic acid site. Conclusion The Sj-specific immune antigen can recognize Sj-specific antigen, and whether the antigen is female sex-specific gene and its value as a candidate gene of Schistosoma japonicum vaccine remains to be further studied.