Evaluation of the transcription level of the protein disulfide isomerase in different stages from Ancylostoma caninum with a real-time PCR assay.
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.
Epe
Christian
C
Institute for Parasitology, University of Veterinary Medicine Hannover, Buenteweg 17, 30559 Hannover, Germany. Christian.Epe@tiho-hannover.de
Behrens
Claudia
C
Strube
Christina
C
Schnieder
Thomas
T
D000722:Ancylostoma / Q000201:enzymology* / Q000254:growth & development* / Q000472:pathogenicity; D000818:Animals; D004285:Dogs; D005260:Female; D005786:Gene Expression Regulation; D015801:Helminth Proteins / Q000235:genetics / Q000378:metabolism; D007814:Larva / Q000201:enzymology / Q000254:growth & development; D008018:Life Cycle Stages; D008297:Male; D016133:Polymerase Chain Reaction / Q000379:methods*; D019704:Protein Disulfide-Isomerases / Q000235:genetics / Q000378:metabolism*; D012337:RNA, Ribosomal, 18S / Q000235:genetics
D015801:Helminth Proteins; D012337:RNA, Ribosomal, 18S; D019704:Protein Disulfide-Isomerases