CAJ | 학술논문

目的：为了寻找血吸虫病新的免疫学诊断候选分子，在大肠杆菌中克隆、表达日本血吸虫（ SjP7）蛋白，建立纯化重组抗原间接ELISA方法（ rSjP7-ELISA）诊断日本血吸虫病。方法设计引物，用PCR法扩增出日本血吸虫 P7样蛋白基因编码基因序列，T载体连接，再将其亚克隆入pET28a（+）表达载体中，经酶切、测序证实正确后，用IPTG对该重组菌诱导表达，SDS-PAGE和Western blot验证表达量和免疫活性。结果 PCR法扩增出一条大小约为423 bp大小的SjP7样蛋白的DNA片断，将其克隆亚表达质粒pET28a（+），测序证实具有完整的ORF。结论日本血吸虫P7蛋白在血吸虫病免疫诊断上具有较大的应用潜能。
목적：위료심조혈흡충병신적면역학진단후선분자，재대장간균중극륭、표체일본혈흡충（ SjP7）단백，건립순화중조항원간접ELISA방법（ rSjP7-ELISA）진단일본혈흡충병。방법설계인물，용PCR법확증출일본혈흡충 P7양단백기인편마기인서렬，T재체련접，재장기아극륭입pET28a（+）표체재체중，경매절、측서증실정학후，용IPTG대해중조균유도표체，SDS-PAGE화Western blot험증표체량화면역활성。결과 PCR법확증출일조대소약위423 bp대소적SjP7양단백적DNA편단，장기극륭아표체질립pET28a（+），측서증실구유완정적ORF。결론일본혈흡충P7단백재혈흡충병면역진단상구유교대적응용잠능。
Objective:To discover new candidate diagnosis molecular for schistosomiasis,the SjP7 protein gene of Schis-tosoma japonicum were cloned and expressed in E. coli BL21. Indirect ELISA(rSjP7-ELISA)was established for schisto-somiasis. Primers were designed and synthesized. Methods:The target gene was amplified from cDNA library by PCR. The product from PCR was cloned into PMD-T vector,and then was subcloned into the expression vector pET28a( +). After induced by IPTG,the cells were collected to analyze by SDS-PAGE and Western blot. Results:For PCR,a specific band of around 423bp was amplified. The gene was subcloned into the expression vector pET28a( +). Conclusion:The rSjP7 has the potential of practical use for the immunodiagnosis of schistosomiasis.