MOLECULAR CHARACTERIZATION OF ESCHERICHIA COLI STRAINS USING PCR-SSCP AND HETERODUPLEX ANALYSIS

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dc.contributor.advisorPurohit, J. H.
dc.contributor.authorAnand, Sanjeev Kumar
dc.date.accessioned2018-06-20T12:31:56Z
dc.date.available2018-06-20T12:31:56Z
dc.date.issued2001
dc.description.abstractThe present study was undertaken with a view to characterize and classify the various strains of E.coli on the basis of SSCP and heteroduplex analysis. DNA from 150 isolates of E.coli, collected earlier from autopsied birds, was extracted using either lysozyme or proteinase K method and V3 region of 168 rRNA gene was amplified by PCR using universal primer designed for that region. The PCR product was subjected to SSCP and heteroduplex analysis on different gel matrices. Five different gel matrices were tested to find out suitability for SSCP and heteroduplex analysis. Among the gels tested 0.5X MDE with 10% glycerol gave the best results. Twenty two different SSCP band patterns were obtained by electrophoresis of ssDNA on non-denaturing gel matrix. The bands of ssDNA in these patterns corresponded to 983 to 379bp equivalent mobility of dsDNA ladder. These patterns contained a minimum of two ssDNA bands to maximum seven. Total 25 bands were found. On heteroduplex analysis keeping the target DNA of Proteus vulgaris as constant, 13 distinct band patterns were observed. The bands of DNA in these patterns corresponded to 900 to 331bp equivalent mobility of dsDNA ladder. These patterns contained a minimum of three to maximum seven heteroduplex bands. The total 17 bands were observed. No relationship could be observed amongst the serotypes, biotypes and SSCP band patterns of E.coli studied. Thus PCR-SSCP and heteroduplex analysis can be used independently for the analysis of the bacterial communities.en_US
dc.identifier.urihttp://krishikosh.egranth.ac.in/handle/1/5810054674
dc.keywordsMOLECULAR CHARACTERIZATION, ESCHERICHIA COLI STRAINS, USING PCR-SSCP, HETERODUPLEX ANALYSISen_US
dc.language.isoenen_US
dc.publisherAAU, Ananden_US
dc.research.problemMOLECULAR CHARACTERIZATION OF ESCHERICHIA COLI STRAINS USING PCR-SSCP AND HETERODUPLEX ANALYSISen_US
dc.subVeterinary Microbiologyen_US
dc.subjectVETERINARY MICROBIOLOGYen_US
dc.subjectCHARACTERIZATIONen_US
dc.themeMOLECULAR CHARACTERIZATION OF ESCHERICHIA COLI STRAINS USING PCR-SSCP AND HETERODUPLEX ANALYSISen_US
dc.these.typeM.V.Sc.en_US
dc.titleMOLECULAR CHARACTERIZATION OF ESCHERICHIA COLI STRAINS USING PCR-SSCP AND HETERODUPLEX ANALYSISen_US
dc.typeThesisen_US
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