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Acta Parasitologica

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Volume 62, Issue 2 (Jun 2017)

Issues

cDNA library construction of two human Demodexspecies

DongLing Niu
  • Department of Pathogen Biology and Immunology, School of Basic Medical Sciences, Xi’an Jiaotong University, Xi’an 710061, China
  • Other articles by this author:
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/ RuiLing Wang
  • Department of Pathogen Biology and Immunology, School of Basic Medical Sciences, Xi’an Jiaotong University, Xi’an 710061, China
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/ YaE Zhao
  • Corresponding author
  • Department of Pathogen Biology and Immunology, School of Basic Medical Sciences, Xi’an Jiaotong University, Xi’an 710061, China
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/ Rui Yang
  • Department of Pathogen Biology and Immunology, School of Basic Medical Sciences, Xi’an Jiaotong University, Xi’an 710061, China
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/ Li Hu
  • Department of Pathogen Biology and Immunology, School of Basic Medical Sciences, Xi’an Jiaotong University, Xi’an 710061, China
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/ YuYang Lei
  • Department of Pathogen Biology and Immunology, School of Basic Medical Sciences, Xi’an Jiaotong University, Xi’an 710061, China
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/ WeiChao Dan
  • Department of Pathogen Biology and Immunology, School of Basic Medical Sciences, Xi’an Jiaotong University, Xi’an 710061, China
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Published Online: 2017-04-18 | DOI: https://doi.org/10.1515/ap-2017-0043

Abstract

The research of Demodex, a type of pathogen causing various dermatoses in animals and human beings, is lacking at RNA level. This study aims at extracting RNA and constructing cDNA library for Demodex. First, P. cuniculiand D. farinaewere mixed to establish homogenization method for RNA extraction. Second, D. folliculorumand D. breviswere collected and preserved in Trizol, which were mixed with D. farinaerespectively to extract RNA. Finally, cDNA library was constructed and its quality was assessed. The results indicated that for D. folliculorum& D. farinae, the recombination rate of cDNA library was 90.67% and the library titer was 7.50 × 104 pfu/ml. 17 of the 59 positive clones were predicted to be of D. folliculorum; For D. brevis& D. farinae, the recombination rate was 90.96% and the library titer was 7.85 x104 pfu/ml. 40 of the 59 positive clones were predicted to be of D. brevis. Further detection by specific primers demonstrated that mtDNA cox1, cox3and ATP6 detected from cDNA libraries had 96.52%-99.73% identities with the corresponding sequences in GenBank. In conclusion, the cDNA libraries constructed for Demodexmixed with D. farinaewere successful and could satisfy the requirements for functional genes detection.

Keywords: Demodex; homogenization method; RNA extraction; cDNA library; quality assessment

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About the article


Received: 2016-11-16

Revised: 2016-12-20

Accepted: 2017-01-19

Published Online: 2017-04-18

Published in Print: 2017-06-01


Citation Information: Acta Parasitologica, ISSN (Online) 1896-1851, ISSN (Print) 1230-2821, DOI: https://doi.org/10.1515/ap-2017-0043.

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© 2017 W. Stefański Institute of Parasitology, PAS. Copyright Clearance Center

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