东方田鼠感染血清免疫筛选日本血吸虫成虫cDNA文库

东方田鼠 (Microtusfortis ,Mf)对日本血吸虫 (Schistosomajaponicum ,Sj)感染具有抗性 .为探讨Mf感染Sj后是否产生针对虫体某些特异抗原分子的免疫应答 ,用Mf感染血清对Sj成虫cDNA文库进行免疫筛选 .经初筛和复筛 ,共筛选出 12个阳性克隆 .这些阳性克隆经辅助噬菌体自动剪切后PCR扩增显示 ,插入的SjcDNA片段大小在 3 0 0bp至 1 8kb之间 ,其中 1 8kb片段 5个 ,1kb片段 1个 ,3 0 0bp片段6个 .经DNA测序分析 ,鉴定出 3个未曾报道过的Sj新基因 ,分别命名为Sj Mf1、Sj Mf2和Sj胞质氨基肽酶 ,并在GenBank登记注册 .结果说明 ,Mf感染血清可识别Sj的特异性抗原分子 ,这些抗原分子的免疫保护作用值得进一步研究 .

Immunoscreening of Schistosoma japonicum Adult Worm cDNA Library by Sera from Infected Microtus fortis

Microtus fortis (Mf) is resistant to Schistosoma japonicum (Sj) infection.To explore if Mf responses to special Sj antigens after infection,Sj adult worm cDNA library was screened with sera from infected Mf.Twelve positive clones were obtained after rescreening.After automatically excised with the helper phage,positive clones were further identified by PCR and DNA sequencing.The size of Sj cDNA fragments in positive clones ranged from 300 bp to 1.8 kb.Five of these fragments were 1.8 kb,one was 1 kb,and six were 300 bp in length.After DNA sequencing,it was discovered that there were three new Sj genes in these clones.These new Sj genes were designated as Sj Mf1,Sj Mf2 and Sj cytosol aminopeptidase and registered in GenBank.The results indicated that the infected Mf serum could recognize Sj specific antigens.It was valuable to the further study immune protection of these cloned molecules.