The differentiation of transcription between tachyzoites and bradyzoites of in vitro cultured Neospora caninum.
Interconversion between tachyzoites and bradyzoites of Neospora caninum plays a pivotal role in transmission of the parasite. Although significant efforts have been made toward understanding the mechanisms that trigger and control stage conversion of the parasite, little is known about this process. We used annealing control primer (ACP)-based polymerase chain reaction (PCR) technique to characterize the differences in transcription between tachyzoites and bradyzoites of N. caninum. The in vitro stage conversion of N. caninum-infected Vero cells was induced by treatment of infected cultures with 70 microM sodium nitroprusside. Subsequently, the gene expression profiles of the tachyzoites and bradyzoites were analyzed through comparison of the level of messenger RNA expression. ACP-based PCR revealed 85 amplicons that were consistently differentially expressed between tachyzoite and bradyzoite stages. Of the 85 differentially expressed transcripts identified, ten were cloned into Topo TA cloning vector, sequenced, and further analyzed by the Basic Local Alignment Search Tool. These differentially expressed transcripts include a combination of known genes and as yet unidentified genes. The present work provides candidate genes for further investigation on molecular basis of stage conversion from tachyzoites to bradyzoites of N. caninum.
Kang
Seung-Won
SW
Department of Bacteria and Parasitology, National Veterinary Research and Quarantine Service (NVRQS), 480 Anyang 6 dong, Anyang City, 420-480, Korea. kangsw@mail.nvrqs.go.kr
Kweon
Chang-Hee
CH
Lee
Eun-Hang
EH
Choe
Se-Eun
SE
Jung
Suk-Chan
SC
Quyen
Dong Van
DV
D000818:Animals; D005786:Gene Expression Regulation / Q000502:physiology*; D018688:Neospora / Q000378:metabolism*; D015800:Protozoan Proteins / Q000235:genetics / Q000378:metabolism; D014158:Transcription, Genetic / Q000502:physiology*
D015800:Protozoan Proteins