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Molecular Cloning and Characterization of Mx2 for Early Pregnancy Diagnosis in Bubalus bubalis

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Proceedings of the National Academy of Sciences, India Section B: Biological Sciences Aims and scope Submit manuscript

Abstract

India is the largest milk producing country in the world. This major achievement is only possible due to the presence of one of the species Bubalus bubalis (Buffalo). Early pregnancy diagnosis in buffalo is one of the major problems related to bovine reproduction system due to silent heat in this animal. Therefore, in the present study, one of the conceptus derived protein myxovirus resistance 2 (Mx2) released during 18–21 days of pregnancy was analyzed by conventional PCR, qPCR and characterized for its application as a biomarker for early pregnancy diagnosis in buffalo. Conventional PCR has shown remarkable amplification in the pregnant animal. Temporal expression of Mx2 revealed its significant increase (P < 0.05) in transcription level during 18–21 days of pregnancy. In silico analysis of protein was done in order to find the suitability of protein as a candidate molecule for early pregnancy diagnostic kits. The functional characterization identifies various motifs present in protein which are responsible for its interaction with other proteins. Physiochemical properties predicted the Mx2 protein nature during in vitro conditions which are required for any of the assays development. However, immunogenic studies revealed that Mx2 is strongly antigenic in nature and can be used for antibody production. In conclusion, Mx2 protein expression from buffalo may be a good indicator of conceptus implantation and can be used in early pregnancy diagnostic kits.

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Acknowledgements

The authors would like to thank veterinarians and non-teaching staffs for collection of samples. They also acknowledge funding support from Council of Scientific and Industrial Research (CSIR), New Delhi for providing fellowship assistance and Rashtriya Krishi Vikas Yojana (RKVY) for the research cost of whole experiment.

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Authors and Affiliations

  1. Department of Animal Biotechnology, College of Veterinary Sciences, Lala Lajpat Rai University of Veterinary and Animal Sciences, Hisar, 125 004, Haryana, India

    Kanisht Batra, Trilok Nanda, Aman Kumar, Rajni Kumari, Vinay Kumar & Sushila Maan

  2. Department of Veterinary Microbiology, College of Veterinary Sciences, Lala Lajpat Rai University of Veterinary and Animal Sciences, Hisar, 125 004, Haryana, India

    Akhil Kumar Gupta

  3. Department of Animal Nutrition, College of Veterinary Sciences, Lala Lajpat Rai University of Veterinary and Animal Sciences, Hisar, 125 004, Haryana, India

    Nancy Sheoran

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  1. Kanisht Batra
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  2. Trilok Nanda
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  3. Aman Kumar
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Correspondence to Kanisht Batra.

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The authors declare that they have no conflict of interest.

Additional information

Significance Statement The characterization study of Mx2 molecule is need of hour for development of early pregnancy diagnostic kit in buffalo at 18–21 days of pregnancy. This characterization will be helpful in prediction of scope of Mx2 as a biomarker in related species.

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Batra, K., Nanda, T., Kumar, A. et al. Molecular Cloning and Characterization of Mx2 for Early Pregnancy Diagnosis in Bubalus bubalis. Proc. Natl. Acad. Sci., India, Sect. B Biol. Sci. 89, 801–814 (2019). https://doi.org/10.1007/s40011-018-0993-x

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  • DOI: https://doi.org/10.1007/s40011-018-0993-x

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