nanos1基因在日本血吸虫不同发育时期的定位  

Location of nanos1 gene in different developmental stages of Schistosoma japonicum

作　　者：华梦晴 邵延靖 刘淼[2] 沈际佳[2] Hua Mengqing;Shao Yanjing;Liu Miao(Dept of Immunology,Anhui Provincial Key Laboratory of Infection and Immunity,Dept of Clinical Laboratory,Bengbu Medical College, Bengbu 233030;Dept of Parasitology, Anhui Medical University, Hefei 230032)

机构地区：[1]蚌埠医学院检验医学院免疫学教研室安徽省感染与免疫重点实验室,蚌埠233030 [2]安徽医科大学基础医学院寄生虫学教研室,合肥230032

出　　处：《安徽医科大学学报》2020年第2期215-221,共7页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:81271865);蚌埠医学院自然科学基金(编号:BYKY1646)。

摘　　要：目的探讨nanos1基因在日本血吸虫的定位以及不同发育时期的表达情况。方法在NCBI上搜索日本血吸虫nanos1基因,PCR扩增、测序并将其全长序列经BLAST软件分析,利用DNAMAN6.0和ClustalX2.0.9进行多重序列比对,利用MAGA 5.05构建进化树进行系统进化分析。体外转录地高辛(DIG)标记的RNA探针,并将其用于日本血吸虫感染后第18、24和42天3个发育时期的雌雄虫整体原位杂交,以确定nanos1基因在日本血吸虫的定位。结果PCR扩增获得日本血吸虫nanos1基因,生物信息学分析结果表明其编码的蛋白与曼氏血吸虫nanos2蛋白同源性最高。体外转录成功的RNA探针用于日本血吸虫整体原位杂交,在感染后第24、42天雌虫的卵巢和卵黄腺以及18天雌虫的卵巢检测到阳性杂交信号,而各个发育阶段的雄虫均未观察到明显阳性杂交信号。结论日本血吸虫nanos1基因高表达于感染后第18天雌虫的卵巢以及第24、42天雌虫的卵巢和卵黄腺。该研究初步揭示了日本血吸虫nanos1基因的表达模式,为研究nanos1基因的功能提供了重要线索。Objective To explore the localization and expression of nanos1 gene in Schistosoma japonicum at different developmental stages.Methods The nanos1 gene of Schistosoma japonicum was searched in NCBI,amplified by PCR and analyzed by BLAST software.DNAMAN 6.0 and ClustalX 2.0.9 were used for multiple sequence alignment,and MAGA 5.05 was used to construct phylogenetic tree for phylogenetic analysis.Digoxin(DIG)labeled RNA probes were transcribed in vitro and used for whole mount in situ hybridization to determine the localization of nanos1 gene in Schistosoma japonicum at 3 developmental stages:18,24 and 42 days.Results The nanos1 gene of Schistosoma japonicum was amplified by PCR.Bioinformatics analysis showed that the highest homology was found between its coding protein and nanos2 protein of Schistosoma mansoni.RNA probes were successfully transcribed in vitro for whole mount in situ hybridization of Schistosoma japonicum at different developmental stages.Positive signals were detected in the ovaries and vitellarium of 24-day-old and 42-day-old females,and in the ovaries of 18-day-old females.No significant positive signal was observed in males at all developmental stages.Conclusion Nanos1 gene is highly expressed in the ovaries and vitellarium of 24-day-old and 42-day-old females,and in the ovaries of 18-day-old females,but no significant positive signal is observed in males at any developmental stages.The expression pattern of nanos1 gene in Schistosoma japonicum is observed successfully,which provided an important clue for studying the function of nanos1 gene.

关 键 词：日本血吸虫 nanos1 进化分析 整体原位杂交 

分 类 号：R383.2[医药卫生—医学寄生虫学]