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Schistosoma japonicum: screening of cercariae cDNA library by specific single-chain antibody against SIEA26-28 ku and immunization experiment of the recombinant plasmids containing the selected genes

机译:日本血吸虫:用针对SIEA26-28 ku的特异性单链抗体筛选尾cDNA文库,并对含有所选基因的重组质粒进行免疫实验

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摘要

To obtain the gene encoding SIEA26-28 ku, which has been proven to be a potential anti-schistosomiasis vaccine candidate, screening Schistosoma japonicum (Sj) cercariae cDNA library with soluble specific single-chain antibody (SIEA26-28 ku-scFv) was performed. A large amount of specific single-chain antibody was harvested through construction of recombinant expression vector pET32a/scFv. The protein was purified and characterized. By using this protein (PET32a-scFv) as a probe, S. japonicum cercariae cDNA library was screened. Two strong positive clones were selected, and their eukaryotic recombinant plasmids were constructed. These genes were named as S. japonicum ribosomal protein S4 (SjRPS4) and S. japonicum ribosomal protein L7 (SjRPL7), respectively. Experiments of mice showed that both SjRPS4 and SjRPL7 DNA vaccines could induce significant immunoprotection. Result of these experiments further proved that the specific single-chain antibody is a very valuable tool in screening of cDNA library to get the corresponding molecules.
机译:为了获得已被证明是潜在的抗血吸虫病疫苗候选基因的编码SIEA26-28 ku的基因,使用可溶性特异性单链抗体(SIEA26-28 ku-scFv)筛选了日本血吸虫尾ScDNA文库。 。通过构建重组表达载体pET32a / scFv,收获了大量特异性单链抗体。纯化蛋白质并鉴定。通过使用该蛋白质(PET32a-scFv)作为探针,筛选了日本血吸虫尾cDNA文库。选择两个强阳性克隆,并构建它们的真核重组质粒。这些基因分别命名为日本血吸虫核糖体蛋白S4(SjRPS4)和日本血吸虫核糖体蛋白L7(SjRPL7)。小鼠实验表明,SjRPS4和SjRPL7 DNA疫苗均可诱导明显的免疫保护。这些实验结果进一步证明,特异性单链抗体是筛选cDNA文库以获得相应分子的非常有价值的工具。

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