摘要
目的 为了寻找新的预防日本血吸虫病的候选疫苗分子。方法 设计合成引物 ,以日本血吸虫成虫基因组DNA为模板 ,用PCR法扩增出丝氨酸蛋白酶 (Sp)基因编码序列 ,其大小约 45 0bp ,将其克隆入pGEM T载体 ,进行序列测定 ,并将Sp基因克隆入真核表达载体 pBKCMV中。 结果 PCR法扩增出SjSp基因编码序列 ,其大小约 45 0bp ,重组质粒 pGEM T Sp和 pBK Sp经双酶切和以质粒为模板进行PCR扩增 ,均可获得一条与PCR产物一致的DNA片段 ,序列测定结果表明具有一个长度为 42 7bp的不完整开放阅读框 ,其与曼氏血吸虫丝氨酸蛋白酶 (SmSp1)具有高度同源性。 结论 本文首次报道了日本积压吸虫丝氨酸蛋白酶 (SjSp)的部分基因编码序列 ,并克隆入了真核表达载体 pBKCMV中 ,为进一步研究提供了条件。
Aim In order to discover new candidate vaccine antigen for schistosomiasis.Methods Primers were designed and synthesized.The partial DNA encoding Sp of S.japonicum was amplified by polymerase chain reaction by using adult worm genomic DNA as template.The product from PCR was cloned into pGEM-T vector and sequenced. Then DNA encoding SjSp was cloned into the eukaryocyte expression vector (pBKCMV).The recombinant vector was identified by restriction analysis and PCR.Results For PCR,a specific band of around 450bp was amplified.The same band was obtained by double restriction of recombinant plasmids and PCR by using recombinant plasmids as template.The nucleotide sequence has an open reading frame of 427bp.Conclusion We firstly reported a partial sequence encoding a novel Schistosima japonicum serine protease.It provided the basis for further study on SjSp expression and its immunological diagnosis and prevention.
出处
《中国人兽共患病杂志》
CSCD
北大核心
2001年第2期25-27,共3页
Chinese Journal of Zoonoses
基金
国家自然科学基金! (编号 ) 3 95 70 64 6)
安徽省自然科学基金资助课题! (编号 97410 0 42 )
