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用抗体探针筛选日本血吸虫成虫cDNA文库

Screening of Schistosoma Japonicum Adult Worm cDNA Library by Antibody Probe

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【作者】 沈定文; 罗金萍; 詹希美; 刘启文;

【Author】 Shen Dingwen et al Department of Parasitology

【机构】 咸宁医学院寄生虫学教研室; 中山医科大学寄生虫学教研室;

【摘要】 本文将重组体噬菌斑的表达抗原转移至硝酸纤维素膜上,用经假筛选法纯化的日本血吸虫感染兔血清(IRS)来检测。结果6×10~5个噬菌斑形成单位(pfu)初筛出23个可能阳性噬菌斑,经再次筛选,其中有8个克隆持续出现阳性。结果表明该免疫筛选能有效地分离单个编码日本血吸虫抗原的克隆,筛选效率高,特异性强,不需同位素标记技术,操作简便,为进一步筛选编码诱导日本血吸虫保护性免疫力的抗原基因打下了基础。

【Abstract】 The antigens produced by recombinant λgt11 plaques were transferred to nitrocellulose filters. Clones encoding given antigens were detected with infected rabbit sera(IRS) which were preabsorbed with NC filters as a catching antibody probe. Twenty-three putative positives were obtained from 6×105 plaque forming units(pfu) in the primary screening and were rescreened a second time,eight of them consistently gave good positive signals in subsequent screenings. The results indicated that the immunoscreening method described here was able to efficiently isolate single cbnes encoding S. japonicum antigens. The method has the advantages of screening libraries efficiently,reliably and specifically. This might open the way to screen genes encoding S. japonicum antigens inducing protective immunity.

  • 【文献出处】 咸宁医学院学报 ,Journal of Xianning Medical College , 编辑部邮箱 ,1995年04期
  • 【分类号】R346
  • 【被引频次】3
  • 【下载频次】25
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