Please use this identifier to cite or link to this item: http://10.1.7.192:80/jspui/handle/123456789/4008
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dc.contributor.authorVaidya, Soumya-
dc.contributor.authorNagar, Nidhi-
dc.contributor.authorPande, Ravi-
dc.contributor.authorZala, Rajpriya-
dc.date.accessioned2013-11-22T09:42:46Z-
dc.date.available2013-11-22T09:42:46Z-
dc.date.issued2013-05-
dc.identifier.urihttp://10.1.7.181:1900/jspui/123456789/4008-
dc.description.abstractLactic acid bacteria (LAB) are considered as generally regarded as safe organism (GRAS). Exploitation of Lactic acid bacteria is advantageous not only in improving the microbial safety of food but also as a probiotic in animals and humans to improve the balance of microflora and to inhibit pathogenic bacteria in the intestinal tract. Screening and isolation of such Lactic acid bacteria from different sources, having probiotics characteristics can be of great importance. In the present study two isolates of lactobacilli, SRN3 and SRN4 from male wistar rat fecal sample were isolated from 36 isolates and screened by various screening methods such as screening in MRS media in presence of Bromo cresol purple dye (BCP) (0.17g/L), MRS+Vancomycin, 2.5 pH adjusted MRS agar plate as well as MRS+0.3% bile (Oxgall) agar plate, sensitivity against various antibiotics and characterization for its probiotic properties such as cell surface hydrophobicity, autoaggregation and coaggregation. Three isolates of lactic acid bacteria, SSR11, SSR14 and SSR16 from fish intestine were isolated from 56 isolates and were screened by various screening methods such as MRS+BCP (0.17g/L), MRS+Vancomycin ,2.5 pH adjusted MRS agar plate as well as MRS+0.3% bile (Oxgall) agar plate, sensitivity against various antibiotics and characterized for its probiotic properties such as acid and bile tolerance, cell surface hydrophobicity, autoaggregation and coaggregation. The antimicrobial activity of the cell free supernatant of all the isolates was determined and found to be effective against different pathogens The crude protein from all the isolates were obtained by ammonium sulphate precipitation and their molecular mass was determined by Sodium Dodecyl Sulphate Poly Acrylamine Gel Electrophoresis. The antimicrobial activity of SRN4 crude protein was found to be of bacteriostatic mode of action against A. hydrophila. Separation and purification of antimicrobial protein of SRN4 was performed by using Superdex 75 (10/300 GL) linked to Reverse Phase-High Performance Liquid Chromatography. The eluted fractions from Size Exclusion Chromatography would be checked for antimicrobial activity and will be purified. 1 The genetic modification of lactobacilli isolates SRN3 and SRN4 is needed to improve the effectiveness of its existing properties and to add new beneficial activities such as its use as a delivery system. Three different strategies have been applied to add new beneficial properties in both the isolates SRN3 and SRN4. The antimicrobial activity of isolates can be increased by incorporation of ColE2 (colicin) gene through genetic alteration. The shuttle vector pRV86 was used for the incorporation ColE2 gene and eletrotransformed into Lactobacilli isolates. For their application as a delivery vehicle and expression system, incorporation of a heterologous signal peptide slpA which provides extracellular secretion properties of the required gene of interest is needed. The secretion vector pSLP111.3 was used to get the slpA signal peptide by corresponding restriction endonucleases and will be incorporated into pRV86 and further transformed into probiotic isolates.Tagging of fluorescent labels by green fluorescent protein (gfp) to the isolates would be a beneficial approach to investigate the colonization potential of probiotics in the gastrointestinal tract after oral administration.en_US
dc.language.isoenen_US
dc.relation.ispartofseriesSDR00185en_US
dc.subjectBiotechnology 2013en_US
dc.subjectProject Report 2013en_US
dc.subjectBiotechnology Project Reporten_US
dc.subjectProject Reporten_US
dc.subject11MBTen_US
dc.subject11MBT008en_US
dc.subject11MBT011en_US
dc.subject11MBT017en_US
dc.subject11MBT020en_US
dc.subjectSDRen_US
dc.subjectSDR00185en_US
dc.titleScreening and Genetic Modification of Antimicrobial Protein Producing Probiotic Lactic Acid Bacteriaen_US
dc.typeDissertationen_US
Appears in Collections:Dissertation, BT

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