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- Title
Actin-depolymerizing factor of second-generation merozoite in <i>Eimeria tenella</i>: clone, prokaryotic expression, and diclazuril-induced mRNA expression.
- Authors
Hong-wei Wang; Fei-qun Xue; Xiao-yang Wang; Feng-kun Yang; Man-man Ban; Rui-xiang Xin; Cong-cong Wang
- Abstract
Abstract  Actin depolymerizing factor (ADF) is an essential actin-binding protein that plays a key role in the control of actin dynamics and actin-based motility processes in intracellular parasites. To determine the effects of diclazuril on ADF gene of second-generation merozoites (mz-ADF) mRNA expression in Eimeria tenella, mz-ADF gene was cloned by RT-PCR from extracted RNA in second-generation merozoite of E. tenella and successfully expressed by pET-28a vector in Escherichia coli BL21(DE3). Results showed that the full length of the cloned cDNA sequence of the mz-ADF gene is 476 bp including an ORF of 375 bp. The sequence has 100% homology with a published sequence of sporozoite stage E. tenella ADF mRNA (GenBank EF195234.1). The recombinant protein was induced to be expressed by 1 mM isopropyl β-d-1-thiogalactopyranoside in vitro. Sodium dodecyl sulfateâpolyacrylamide gel electrophoresis analysis showed that 16.99 kDa fusion protein existed in solvable form. Compared with the infected/control group, mz-ADF mRNA expression level was downregulated by 63.86% in the infected/treatment group with the treatment of diclazuril. In conclusion, the data presented here indicate that mz-ADF gene participates in an important role in the invasion host of E. tenella. Downregulation of mz-ADF mRNA expression enrich the mechanism study of diclazuril on E. tenella.
- Subjects
EIMERIA; MICROFILAMENT proteins; PROKARYOTES; MESSENGER RNA; ACTOMYOSIN; ANTIPARASITIC agents; PHARMACODYNAMICS; REVERSE transcriptase polymerase chain reaction
- Publication
Parasitology Research, 2010, Vol 106, Issue 3, p571
- ISSN
0932-0113
- Publication type
Article