Epe Christian, Behrens Claudia, Strube Christina, Schnieder Thomas
Institute for Parasitology, University of Veterinary Medicine Hannover, Buenteweg 17, 30559 Hannover, Germany.
Parasitol Res. 2007 Nov;101(6):1589-95. doi: 10.1007/s00436-007-0679-4. Epub 2007 Aug 28.
The protein disulfide isomerase (PDI) is a ubiquitous protein, which contributes in building disulfide bridges. In the work presented here, the expression of the PDI in different stages of the canine hookworm Ancylostoma caninum was investigated. Third-stage larvae (L3), adults, as well as serum-stimulated and hypobiotic L3 were used. For quantification of the PDI gene transcription, a real-time PCR was used establishing a hybridization probe (TaqMantrade mark probes) for detection of PDI copy numbers in different populations. 18S ribosomal ribonucleic acid (rRNA) was used as a housekeeping gene for normalization. The results show differences in the transcription level of the investigated A. caninum populations: The serum-stimulated larvae representing the switch to parasitism showed the highest PDI expression. The hypobiotic larvae representing a resting stage showed the lowest expression level. Male adults showed an elevated expression compared to female adult worms. The L3 expression level was just below the serum-stimulated population. This work confirms the upregulated gene expression of PDI during host penetration and invasion.
蛋白质二硫键异构酶（PDI）是一种普遍存在的蛋白质，它有助于形成二硫键。在本文所述的研究中，对犬钩虫（犬弓首线虫）不同发育阶段的PDI表达进行了研究。使用了第三期幼虫（L3）、成虫以及经血清刺激的滞育L3。为了定量PDI基因转录，采用实时PCR技术，建立了一种杂交探针（TaqMan™探针）用于检测不同虫群中的PDI拷贝数。18S核糖体核糖核酸（rRNA）用作管家基因进行标准化。结果显示，所研究的犬弓首线虫不同虫群的转录水平存在差异：代表向寄生状态转变的经血清刺激的幼虫PDI表达最高。代表静止阶段的滞育幼虫表达水平最低。雄性成虫的表达高于雌性成虫。L3的表达水平略低于经血清刺激的虫群。这项研究证实了在宿主穿透和入侵过程中PDI基因表达上调。
