Department of Parasitology, Xiangya Medical School, Central South University, 110 Xiangya Road, Changsha, 410078, People's Republic of China.
Parasitol Res. 2010 Jun;107(1):127-34. doi: 10.1007/s00436-010-1846-6. Epub 2010 Apr 13.
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.
为了获得已被证明是一种有潜力的抗血吸虫病疫苗候选物的 SIEA26-28 ku 基因,我们用可溶性特异性单链抗体(SIEA26-28 ku-scFv)筛选日本血吸虫(Sj)尾蚴 cDNA 文库。通过构建重组表达载体 pET32a/scFv,大量特异性单链抗体被收获。该蛋白经过纯化和鉴定。利用这种蛋白(PET32a-scFv)作为探针,筛选日本血吸虫尾蚴 cDNA 文库。选择了两个强阳性克隆,并构建了它们的真核重组质粒。这些基因分别被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 疫苗均能诱导显著的免疫保护。这些实验的结果进一步证明,特异性单链抗体是筛选 cDNA 文库获得相应分子的非常有价值的工具。