Actin-depolymerizing factor of second-generation merozoite in Eimeria tenella: clone, prokaryotic expression, and diclazuril-induced mRNA expression

作者：Zhou, Bian-hua; Wang, Hong-wei; Xue, Fei-qun^*; Wang, Xiao-yang; Yang, Feng-kun; Ban, Man-man; Xin, Rui-xiang; Wang, Cong-cong

来源：Parasitology Research, 2010, 106(3): 571-576.

DOI：10.1007/s00436-009-1699-z

• DEVELOPMENTAL STAGES
• CELL POLARIZATION
• FACTOR COFILIN
• INVIVO ACTION
• ADF/COFILIN
• MECHANISM
• INVASION
• DYNAMICS
• CLINACOX

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 beta-d-1-thiogalactopyranoside in vitro. Sodium dodecyl sulfate-polyacrylamide 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.

• 出版日期2010-2
• 单位河南大学; 中国农业科学院上海兽医研究所