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阳性水牛、黄牛IgG筛选日本血吸虫成虫噬菌体展示cDNA文库
  • ISSN号:1674-6422
  • 期刊名称:《中国动物传染病学报》
  • 时间:0
  • 分类:S852.735[农业科学—基础兽医学;农业科学—兽医学;农业科学—畜牧兽医]
  • 作者机构:中国农业科学院上海兽医研究所国家防治动物血吸虫病专业实验室农业部动物寄生虫学重点开放实验室,上海200241
  • 相关基金:公益性行业(农业)科研专项经费(201303037); 农业部动物疫情监测与防治
中文摘要:

为寻找有效的家畜日本血吸虫病诊断抗原,从感染日本血吸虫的水牛、黄牛血清中纯化IgG,以亲和淘洗方法筛选日本血吸虫成虫(44日龄)噬菌体展示c DNA文库。经每种动物IgG三轮(总计六轮)筛选后,随机挑取53个阳性噬菌体克隆进行序列测定及生物信息学分析。获得了9个EST序列,其中7个EST序列与已知基因或表达序列标签同源,2个EST序列与已知基因或表达序列标签均无同源性,以噬菌体展示载体的10B衣壳蛋白开放阅读框(open reading frame,ORF)为依据,对上述7个与Gen Bank数据库资料同源的EST进行对码分析,结果 7个EST序列能和噬菌体展示载体阅读框对码,为有效展示序列。其中3个EST具有较高的重复度,在51个有效克隆中,25个为HSP70 homolog基因,15个为Keratin9基因有重复,6个为Ornithine--oxo-acid transaminase基因。研究结果显示,HSP70 homolog基因、Keratin9基因和Ornithine--oxo-acid transaminase基因作为诊断家畜血吸虫病候选抗原基因,值得进一步开展克隆、表达和诊断效果评估方面的研究。

英文摘要:

In order to find the effective diagnostic antigen for S.japonicum infection in domestic animals, IgG preparations were purified from serum samples of infected buffalo and cattle and used to screen a phage display cDNA library from 44-day-old worms. After a total of six rounds of biopanning, 53 positive clones were randomly picked and sequenced and 9 ESTs including 2 unnamed ESTs were obtained. With open reading frame (ORF) of 10B capsid protein in phage display vector as the basis, 7 ESTs were determined to be effective display sequences. Among the 7 effectively displayed ESTs, 3 ESTs had higher multiplicity. Among the 51 positive clones sequenced, 25 clones were HSP70 homolog, 15 clones were Keratin 9 and 6 clones were Ornithine-oxo-acid transaminase. These results obtained here suggested that the genes encoding HSP70 homolog, Keratin 9 and Ornithine-oxo-acid transaminase might be relevant to the candidate diagnostic antigens for detection of S.japonicum infection in domestic animals.

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期刊信息
  • 《中国动物传染病学报》
  • 中国科技核心期刊
  • 主管单位:中国农业部
  • 主办单位:中国农业科学院上海兽医研究所
  • 主编:童光志
  • 地址:上海市闵行区紫月路518号
  • 邮编:200241
  • 邮箱:bianjibu@shrri.ac.cn
  • 电话:021-34293142
  • 国际标准刊号:ISSN:1674-6422
  • 国内统一刊号:ISSN:31-2031/S
  • 邮发代号:4-748
  • 获奖情况:
  • 第二届全国畜牧兽医优秀期刊二等奖,首届《CAJ-CD规范》执行优秀奖
  • 国内外数据库收录:
  • 美国化学文摘(网络版),中国中国科技核心期刊,中国北大核心期刊(2014版)
  • 被引量:1517