代表差异性分析方法分离日本血吸虫雌虫特异性序列

摘要

目的　分离日本血吸虫雌虫特异性序列，为研制抗生殖疫苗打下基础。方法　应用代表差异性分析方法　利用雄虫基因扣除雌虫基因，从而获得雌虫特异性序列，约562bp。结果　以该序列为探针与S0uthem转印的酶切后的雌、雄虫基因组DNA及检测子和驱动子杂交，免疫学检测，显示只有检测子(tester)泳道有杂交区带，驱动子(driver)区带没有此带。结论　初步说明所获序列为雌虫特异性序列。%Aim To isolate the female-specific sequence of Schistosomajaponicum S. J. in order to provide a basis for researching vaccines of anti-fecundity. Methods Representational difference analysis (RDA) was used to obtain a female-specific sequence of S.j. by the male S.j. genomic DNA substractive hybridization with the female. The fragment was about 562bp. Results This sequence was labeled as Dig probe by PCR technique to hybridize with the restriction fragments of each sex and the tester as well as the driver on the nylon membranes.The antiboly of Dig、 NBT、 and X-phosphate solution were utilized to immunoassay the result of hybridization. The result showed that the probe can hybridize to the tester but can not hybridize to the driver. Conclusion The result suggested that the fragment of approx. 562bp was the female-specific sequence of Schistosoma japonicum.