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International Journal of Current Microbiology and Applied Sciences (IJCMAS)
IJCMAS is now DOI (CrossRef) registered Research Journal. The DOIs are assigned to all published IJCMAS Articles.
Index Copernicus ICI Journals Master List 2019 - IJCMAS--ICV 2019: 96.39 For more details click here
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National Academy of Agricultural Sciences (NAAS)
NAAS Score: *5.38 (2020)
[Effective from January 1, 2020]
For more details click here

ICV 2019: 96.39
Index Copernicus ICI Journals Master List 2019 - IJCMAS--ICV 2019: 96.39
For more details click here

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Original Research Articles

PRINT ISSN : 2319-7692
Online ISSN : 2319-7706
Issues : 12 per year
Publisher : Excellent Publishers
Email : editorijcmas@gmail.com / submit@ijcmas.com
Editor-in-chief: Dr.M.Prakash
Index Copernicus ICV 2018: 95.39
NAAS RATING 2020: 5.38

Int.J.Curr.Microbiol.App.Sci.2020.9(12): 476-487
DOI: https://doi.org/10.20546/ijcmas.2020.912.058


Molecular Cloning of Pyruvate Formate Lyase Gene of Staphylococcus aureus Associated with Bovine Mastitis
Varra Manasa1*, T. Venkata Sai Kumar2, T. Prasada Rao2, K. Aswani Kumar3, J. Thanislass4 and V. Girish Kumar5
1Department of VBC, Veterinary College, Hebbal, KVAFSU, India
2Department of VBC, C.V.Sc, PDTR, SVVU, India
3Department of VBC, N.T.R C.V.Sc, Gannavaram, SVVU, India
4Department of VBC, RIVER, Puducherry, India
5Department of VBC, Veterinary College, Hebbal, KVAFSU, India
*Corresponding author
Abstract:

Puruvate formate lyase (PFL) of Staphylococcus aureus is an important protein which gets expressed mainly as an adaptive response for their survival in the host mammary tissue by way of producing biofilm. This adaptive mechanism of S. aureus mainly contributes to sub-clinical, chronic bovine mastitis which often becomes challenging to treat with conventional antibiotics and thereby the prevention of further spread of infection in the herd becomes difficult. The present preliminary study is an attempt to clone and characterize 609 bp antigenic site of pyruvate formate lyase gene of S. aureus associated with bovine mastitis. The partial PFL gene cloned using TA cloning vector showed 98-99% identity with the reference sequence available in the Genbank (MK695632). The cloned insert with overhangs released by restriction enzyme digestion can be used to obtain the antigenic protein by using pET 32a+ expression vector system. The antibodies to pyruvate formate lyase were obtained in the serum of cows associated with S. aureus bovine mastitis which was as an important finding in Serological Proteome Analysis (SERPA). Therefore the protein expressed from the cloned partial PFL gene can be used for the development of immunoassay kit for early diagnosis and also as a vaccine candidate for the control and prevention of S. aureus associated bovine mastitis.


Keywords: Staphylococcus aureus, Bovine mastitis, Pyruvate formate lyase gene (PFL gene), Cloning, Immunogenic protein
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How to cite this article:

Varra Manasa, T. Venkata Sai Kumar, T. Prasada Rao, K. Aswani Kumar, J. Thanislass and Girish Kumar, V. 2020. Molecular Cloning of Pyruvate Formate Lyase Gene of Staphylococcus aureus Associated with Bovine Mastitis.Int.J.Curr.Microbiol.App.Sci. 9(12): 476-487. doi: https://doi.org/10.20546/ijcmas.2020.912.058