旋毛虫抗原基因Ts88的克隆及鉴定

Cloning and identification of a novel antigenic gene(Ts88) of Trichinella spiralis

目的 获取旋毛虫具有抗原性的新基因。方法 应用旋毛虫免疫血清和人工感染血清对旋毛虫成虫cDNA文库约 3× 10 5重组噬菌斑进行筛选。基因克隆、DNA测序及 5′ -RACE获cDNA全长 ;采用ESEE、DNAstar软件及核苷酸序列数据库 (GenBank)进行cDNA序列分析。结果与结论 免疫筛选成虫cDNA文库 ,获得 3个阳性克隆。其中的Ts88克隆cDNA全长为 196 6bp ,编码 4 84个氨基酸 ,含有一个PWWP功能结构域及多个抗原表位。Ts88cDNA是一个尚未见报道的旋毛虫抗原新基因。

Objective To get a novel antigenic gene of Trichinella spiralis (T.s) Methods 3×105 recombinant plagues of adult cDNA library of T.s were screened using the sera from immunized and infected rabbits.The full length of cDNA sequence was obtained by cloning,DNA sequencing and 5′-RACE technique.The cDNA sequence was analyzed by ESEE,DNAstar softwares and GenBank database.Results and Conclusion Three positive clones were identified by immunoscreening.One of them Ts88 cDNA contains 1966bp,which encodes 484 amino acids.A PWWP domain and some possible antigen epitopes were predicated within the amino acids.Ts88 cDNA is a novel antigenic gene of T.s.