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Molecular characterization of an actin depolymerizing factor from Cryptocaryon irritans.
MedLine Citation:
PMID:  23286243     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
SUMMARY Actin depolymerizing factors regulate actin dynamics involved in cellular processes such as morphogenesis, motility, development and infection. Here, a novel actin depolymerizing factor gene (CiADF 2 ) was cloned from the cDNA library of Cryptocaryon irritans, a parasitic ciliate causing cryptocaryonosis. The full-length cDNA of CiADF 2 was 531 bp. Its open reading frame (ORF) was 417 bp, encoding a polypeptide of 138 aa with typical features of the ADF/cofilin family. Reverse transcription-PCR suggested that CiADF 2 is expressed in all stages of the life cycle. After site-directed mutagenesis of a non-universal genetic code, the ORF was subcloned in Escherichia coli. The bacteria were induced with the addition of isopropylthio-β-D-galactoside to express a fusion protein of recombinant CiADF2 (rCiADF2) with glutathione S transferase. Sodium dodecyl sulfate polyacrylamide gel electrophoresis and Western blot confirmed the predicted molecular mass of rCiADF2 of 16·2 kDa. A mouse antibody against rCiADF2 recognized native CiADF2, and rCiADF2 reacted with mouse antisera against C. irritans trophonts. CiADF2 was abundant in the plasma around cytostomes, suggesting that CiADF2 is involved in ciliate movement. Moreover, rCiADF2 showed F-actin binding and depolymerizing activity. This study will help to clarify the pathogenic biology of the parasite and develop effective control measures for cryptocaryonosis.
Authors:
Xiaohong Huang; Yang Xu; Guowei Guo; Qianqian Lin; Zhongfeng Ye; Liping Yuan; Zhiyu Sun; Wei Ni
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Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2013-1-4
Journal Detail:
Title:  Parasitology     Volume:  -     ISSN:  1469-8161     ISO Abbreviation:  Parasitology     Publication Date:  2013 Jan 
Date Detail:
Created Date:  2013-1-4     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0401121     Medline TA:  Parasitology     Country:  -    
Other Details:
Languages:  ENG     Pagination:  1-8     Citation Subset:  -    
Affiliation:
Fujian Provincial Key Laboratory of Developmental Biology and Neuroscience, College of Life Science, Fujian Normal University, Fuzhou 350108, Fujian, China.
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