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目的 克隆并鉴定日本血吸虫酪氨酸羟化酶(TH)基因,探讨其他信号蛋白在信号传导中对TH的调节及其之间的分子作用机制,从而为血吸虫新型疫苗的设计和药物开发开辟新的途径。 方法 以曼氏血吸虫的 TH cDNA为模板设计引物,以日本血吸虫成虫mRNA为模板逆转录合成cDNA链,将扩增出的日本血吸虫TH蛋白编码基因序列,克隆入 pGEM T easy载体,用单酶双切法和DNA测序进行鉴定,并与曼氏血吸虫的TH基因序列进行同源性比较。 结果 自日本血吸虫RNA经逆转录得到一长度为 480 bp的 DNA片段,经测序与曼氏血吸虫 TH的同源性为 87%。结论 成功扩增出日本血吸虫TH的中间编码区域,为进一步扩增日本血吸虫TH基因全长以及后续实验奠定了基础。
Objective To clone and identify the gene of tyrosine hydroxylase (TH) of Schistosoma japonicum and investigate the regulation of TH signaling and the molecular mechanism of its interaction with other signaling proteins, thus to open up a new vaccine design and drug development for schistosomiasis Way. Methods TH cDNA of Schistosoma mansoni was used as a template to design a primer for reverse transcription of adult Schistosoma japonicum mRNA as a template. The gene encoding TH protein of S. japonicum was amplified and cloned into pGEM T easy vector. DNA sequencing and sequencing, and compared with the sequence of TH gene of Schistosoma mansoni. Results A DNA fragment of 480 bp in length was obtained by reverse transcription of Schistosoma japonicum RNA. The sequence was 87% identical to TH of Schistosoma mansoni. Conclusion The intermediate coding region of TH from Schistosoma japonicum was successfully amplified, which laid the foundation for further amplification of TH gene of S. japonicum and subsequent experiments.