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Acta Parasitologica

, Volume 64, Issue 2, pp 251–256 | Cite as

The Construction of Full-Length cDNA Library for Otodectes cynotis

  • Li Hu
  • Ya-e ZhaoEmail author
  • Dong-ling Niu
  • Rui Yang
  • Ji-hui Zeng
Original Paper
  • 26 Downloads

Abstract

Background

Otodectes cynotis (Hering, 1838) is the pathogen of otodectic mange distributed worldwide. The mite mainly infests carnivores and, sometimes, humans. However, due to the lack of cDNA library, research on its pathogenesis has been challenging.

Methods

To solve this problem, the present study first sampled O. cynotis mites from an infested cat from Xi’an, China, for RNA extraction. Then, the full-length cDNA library was constructed using the SMART technique. Finally, positive clones > 500 bp and Hsc70-5 gene fragment specifically amplified from the cDNA library were sequenced and analyzed to verify the library’s reliability.

Results

Results showed that RNA extracted from 300 mites had good quality with a concentration of 149 ng/μl and OD260/OD280 of 1.99. The library satisfied the quality standard of a good library with a titer of 5.02 × 105 PFU/ml and a combination rate of 97.61%. In addition, clone 4 and Hsc70-5 showed 98.38% and 99.72% identity with Ef1-α and Hsc70-5 gene sequences of O. cynotis in GenBank, respectively.

Conclusion

The cDNA library of O. cynotis constructed here was successful and reliable, creating the basis for research on RNA sequencing and functional genes of O. cynotis.

Keywords

Otodectic mange cDNA library Quality evaluation Specific primer detection 

Notes

Acknowledgements

This work was supported by the National Natural Science Foundation of China (Nos. 81471972; 81271856).

Compliance with Ethical Standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

11686_2019_34_MOESM1_ESM.docx (21 kb)
Supplementary material 1 (DOCX 21 kb)
11686_2019_34_MOESM2_ESM.jpg (393 kb)
Supplementary Fig. 1. Sequence analysis of Ef1-α gene. A. Sequence alignments of Otodectes, B. ML tree across Psoroptidia. (JPEG 392 kb)
11686_2019_34_MOESM3_ESM.jpg (294 kb)
Supplementary Fig. 2. Sequence analysis of Hsc70-5 gene. A. Sequence alignments of Otodectes, B. ML tree across Psoroptidia. (JPEG 294 kb)

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Copyright information

© Witold Stefański Institute of Parasitology, Polish Academy of Sciences 2019

Authors and Affiliations

  • Li Hu
    • 1
  • Ya-e Zhao
    • 1
    Email author
  • Dong-ling Niu
    • 1
  • Rui Yang
    • 1
  • Ji-hui Zeng
    • 2
  1. 1.Department of Pathogenic Biology and Immunology, School of Basic Medical SciencesXi’an Jiaotong UniversityXi’anChina
  2. 2.Xi’an PET Animal HospitalXi’anChina

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