Identification of serotype specific dna marker for salmonella typhimurium by rapd-pcr method

Download

index.pdf

Date

2004

Author

Aksoy, Ceren

Metadata

Show full item record

Item Usage Stats

224
views

103
downloads

This study was performed for the identification of specific DNA marker using RAPD-PCR (Random Amplified Polymorphic DNA) method for serotype Salmonella Typhimurium which is one of the most prevalent serotype causing food poisining all over the world. The Primer 3 (RAPD 9.1), 5̕-CGT GCA CGC-3ء, was used in RAPD-PCR with 35 different Salmonella isolates. 12 of them were serotype Salmonella Typhimurium and 23 of them were belonging to other six different serotypes. Accordingly, two different 300 bp and 700 bp sized amplification products were obtained from the 12 different Salmonella Typhimurium isolates. On the other hand, other 23 Salmonella isolates of six different serotypes gave only 300 bp amplification band while 700 bp amplification band was not observed with Primer 3 (RAPD 9.1). After the discovery of 700 bp fragmentwhich was specific for S. Typhimurium, it was decided to sequence. The 700 bp band was ligated onto the vector pUC 19 to sequence. The cloninig operation gave positive results by the formation of blue and white colonies, but plasmid isolation process from white colonies containing the ligated vector was not achieved. Therefore, sequencing of the 700 bp fragment together with plasmid DNA could not be completed. However it wil be sent to USA for sequencing. According to these results, it was discovered that 700 bp amplification product was found as a specific polymorphic region for Salmonella Typhimurium after RAPD application on genomic DNA and this band can be used as a specific marker for detection and identification of Salmonella Typhimurium.

Subject Keywords

Bacteria.

URI

http://etd.lib.metu.edu.tr/upload/12605441/index.pdf
https://hdl.handle.net/11511/14405

Collections

Graduate School of Natural and Applied Sciences, Thesis

Suggestions

OpenMETU
Core

Characterization and purification of a bacteriocin produced by Leuconostoc mesenteroides subsp. cremoris Dündar, Halil; Bozoğlu, Faruk; Department of Biotechnology (2006) In this study, a new bacteriocin isolated from a Leuconostoc mesenteroides subsp. cremoris strain was purified to homogeneity by pH mediated cell adsorption-desorption, solid phase extraction with Amberlite XAD-16, cation-exchange chromatography and hydrophobic interaction chromatography. The purification resulted in an electrophoretically pure protein that was smaller than 6 kDa as judged by SDS-PAGE. The bacteriocin was found to be very hydrophobic and cationic and could be adsorbed by synthetic calcium s...
Effect of traditional starter cultures on quality of cheese Kayagil, Fulya; Gürakan, Candan; Department of Biotechnology (2006) In this study, the physico-chemical chances occurring in white cheese and possible effects of starter culture combinations to the ripening period during 30 days storage examinated. A total of thirty six lactic acid bacteria were isolated from a cheese made witohout using starter culture. For identification gram staining, catalase, gas production and coagulation tests were performed. For determination of species API50 CH (BioMérieux) and partial 16S rDNA gene sequence analysis were used. Four cheese were pro...
Biochemical and genetic characterization of halobacterium salinarium strain isolated from Tuz Lake in central Anatolia Çakıcı, Özgür; Yücel, Ayşe Meral; Department of Biochemistry (2004) In this study, a halophilic archaea Halobacterium salinarium TG13 which is isolated from Tuz Lake in Central Anatolia was characterized biochemically and genetically. Halobacterium salinarium DSM3754 and Halobacterium salinarium S9 strains were used as a reference strain through the experiments. In biochemical characterization; total protein profiles of strains was compared by using 1D SDS PAGE. Total protein profile of the isolated strain has shown differences. The SDS-PAGE profile of the purified purple m...
IDENTIFICATION OF LACTOBACILLUS STRAINS FROM TURKISH-STYLE DRY FERMENTED SAUSAGES Gültekin, Güzin Candan; WEISS, N (1995-01-01) A total of 31 strains of facultatively anaerobic, non-motile, Gram-positive, rod-shaped bacteria were isolated from different Turkish dry sausage (soudjouk) samples. They were determined as belonging to the genus Lactobacillus and subjected to a taxonomic study. The strains were divided into seven groups based on sugar fermentation pattern and other physiological and morphological characteristics. A variety of biochemical and genetic characteristics were also examined for species differentation of these iso...
Behaviour and control of listeria innocua during manufacture and storage of Turkish white cheese Öztürkoğlu, Şebnem; Gürakan, Candan; Department of Food Engineering (2004) Growth and survival of L. innocua and TAC in artificially inoculated Turkish White Cheese during manufacturing and storage periods, with respect to different level of contamination of L. innocua were investigated. Cheese products were manufactured by the short-set procedure in pilot-plant-sized vats, as in AOÇ dairy factory. Pasteurized cow̕s milk was inoculated with L. innocua for obtaining the initial loads of 3.84 and 7.12 log CFU/ml. Bacterial load of inoculated milk, whey, post-ripened curd and post-sa...

Citation Formats

C. Aksoy, “Identification of serotype specific dna marker for salmonella typhimurium by rapd-pcr method,” M.S. - Master of Science, Middle East Technical University, 2004.