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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 2018 - IJCMAS--ICV 2018: 95.39 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 2018: 95.39
Index Copernicus ICI Journals Master List 2018 - IJCMAS--ICV 2018: 95.39
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.2018.7(7): 2827-2835
DOI: https://doi.org/10.20546/ijcmas.2018.707.331


Influence of Growth Stages and Nutrient Media on Production of Membrane Vesicles of Clostridium perfringens Type A and their Molecular Characterization
Deuri Nirab Kumar1*, Sharma Kr. Rajiv2, Borah Probodh1, Hazarika Parishmita2, Sonowal Joyshikh1, Borguhain Indrani1 and Pathak Bedanta1
1Department of Animal Biotechnology
2Department of Veterinary Microbiology,College of Veterinary Science, Assam Agricultural University, Khanapara, Guwahati-781022, India
*Corresponding author
Abstract:

All Gram-negative bacteria and a few gram-positive bacteria shed membrane vesicles in suitable medium and environmental condition. The present study was undertaken to determine the influence of stages of growth and nutrient media on production of membrane vesicles (MVs) of Clostridium perfringens type A (C. perfringens type A), and molecular characterization of the same. An isolate of C. perfringens belonging to type A was selected and grown in Trypticase Peptone Glucose broth (TPGB) and Robertson's Cooked Meat broth (RCMB) media. The MVs were extracted at 4, 8, 12 and 24 hours of growth. The protein profile based on SDS-PAGE revealed the appearance of six bands at 4 and 8 hrs of growth, with one prominent band and nine bands at 12 hrs of growth on TPGB. No bands were observed in MVs extracted at 24 hrs of growth in TPGB. The molecular weight of the protein bands ranged from 43.3 kDa to 75.2 kDa. The MVs extracted from RCMB medium revealed the appearance of six protein bands at 4 and 8 hrs and nine bands at12 hrs of growth with six prominent bands. The 24 hrs growth culture in RCMB also revealed no bands in SDS-PAGE. The RCMB protein bands also ranged from 43.3 kDa to 75.2 kDa. The DNA content of MVs was demonstrated by PCR and presence of DNA 16S rRNA and cpa (alpha toxin) genes in MVs was confirmed. Agarose gel electrophoresis revealed two distinct bands of 795bps and 324 bps of 16S rRNA and cpa (alpha toxin) genes, respectively.


Keywords: Clostridium perfringens, Membrane Vesicles (MVs), SDS-PAGE, PCR
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How to cite this article:

Deuri Nirab Kumar, Sharma Kr. Rajiv, Borah Probodh, Hazarika Parishmita, Sonowal Joyshikh, Borguhain Indrani and Pathak Bedanta. 2018. Influence of Growth Stages and Nutrient Media on Production of Membrane Vesicles of Clostridium perfringens Type A and their Molecular Characterization.Int.J.Curr.Microbiol.App.Sci. 7(7): 2827-2835. doi: https://doi.org/10.20546/ijcmas.2018.707.331