National Academy of Agricultural Sciences (NAAS)
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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 |
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.
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