日本血吸虫重组蛋白TMEM66P的表达和多克隆抗体的制备

Prokaryotic Expression of Recombinant Protein TMEM66P of Schistosoma Japonicum and Preparation of Antiserum

血吸虫病是由血吸虫尾蚴感染引起的一种人畜共患寄生虫病,对宿主造成严重病理性损害,是WHO确定的六大重点热带病之一。血吸虫体被直接与宿主的免疫系统相接触,是宿主与虫体信息交流的重要界面,位于虫体体被跨膜蛋白的功能研究尤显重要。本文对日本血吸虫膜蛋白(TEME66P)的cDNA进行表达和抗血清的制备。首先,利用生物信息学分析TMEM66P跨膜结构,并构建系统进化树,构建重组原核表达质粒并表达纯化重组蛋白,制备抗血清,通过Western blot对抗血清的特异性进行检测。研究表明TMEM66P存在跨膜区,其氨基酸序列与曼氏血吸虫同源性为67.79%,抗血清能够识别出全虫蛋白中TMEM66P,具有良好的特异性。本研究为进一步探索TMEM66P在日本血吸虫中的功能奠定了初步基础。

Schistosomiasis is a zoonotic parasitic disease caused by cercariae infection,causing severe pathological damages to the hosts and one of the six major tropical diseases listed by WHO.The transmembrane protein locates on the tegument of schistosomes where they directly contact with the host's immune system and may play an important role in the signal communication between the host and parasite.In this paper,we reported prokaryotic expression of the cDNA of the transmembrane protein 66 precursor(TEME66P)of Schistosoma japonicum and preparation of antiserum.The fi rst step was to analyze the transmembrane domain of TMEM66P by using bioinformatics software,then to construct a phylogenetic tree and plasmid for expressing the recombinant protein.Next,the recombinant protein was purifi ed and used to prepare the antiserum.The results showed that TMEM66P had a transmembrane region,of which amino acid sequence was 67.79%homologous to S.mansoni.In addition,the antiserum specifi cally reacted with TMEM66P from whole worm's protein.Our preliminary study laid the foundation for further exploring the function of TMEM66P in S.japonicum.