摘要
Objective： To clone and construct the recombinant plasmid containing ATP synthase lipid-binding protein-like protein gene of Schistosomajaponicum,（SjAslp） and transfer it into mammalian cells to express the objective protein. Methods： By polymerase chain reaction （PCR） technique, SjAslp was amplified from the constructed recombinant plasmid pBCSK＋/SjAslp, and inserted into cloning vector pUCm-T. Then, SjAslp was subcloned into an eukaryotic expression vector pcDNA3.1（＋）. After identifying it by PCR, restrictive enzymes digestion and DNA sequencing, the recombinant plasmid was transfected into HeLa cells using electroporation, and the expression of the recombinant protein was analyzed by immunocytochemical assay. Results： The specific gene fragment of 558 bp was successfully amplified. The DNA vaccine of SjAslp was successfully constructed. Immunocytochemical assay showed that SjAslp was expressed in the cytoplasm of HeLa cells. Conclusion： SjAslp gene can be expressed in eukaryotic system, which lays the foundation for development of the SjAslp DNA vaccine against schitosomiasis.
克隆并且构造包含 ATP synthase 类脂化合物绑定的 recombinant plasmid 的目的血吸虫 japonicum 的像蛋白质的蛋白质基因，(SjAslp ) 并且把它变成哺乳动物的房间表示客观蛋白质。由聚合酶链反应(PCR ) 的方法技术， SjAslp 从构造 recombinant plasmid pBCSK+/SjAslp 被放大，并且插入了到克隆向量 pUCm-T。然后， SjAslp 是进真核细胞的表示向量 pcDNA3.1 (+) 的 subcloned。在由定序的 PCR，限制的酶消化和 DNA 识别它以后， recombinant plasmid 是进用 recombinant 蛋白质的 electroporation，和表示的 HeLa 房间的 transfected 被 immunocytochemical 分析试金。结果 558 bp 的特定的基因碎片成功地被放大。SjAslp 的 DNA 疫苗成功地被构造。Immunocytochemical 试金证明 SjAslp 在 HeLa 房间的细胞质被表示。结论 SjAslp 基因能在真核细胞的系统被表示，它对 schitosomiasis 为 SjAslp DNA 疫苗的开发打基础。
基金
the Scientific Research Foundation of Education Committee of Hunan (07C713 and 07C708)
