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Ahani Azari, Ania, Zahraei Salehi, Taghi, Nayeri Fasaei, Bahar, Madadgar, Omid, Alebouyeh, Masoud. (1393). Construction of a recombinant vector for site-directed mutagenesis in Salmonella typhimurium. , 8(3), 187-192. doi: 10.22059/ijvm.2014.51890
Ania Ahani Azari; Taghi Zahraei Salehi; Bahar Nayeri Fasaei; Omid Madadgar; Masoud Alebouyeh. "Construction of a recombinant vector for site-directed mutagenesis in Salmonella typhimurium". , 8, 3, 1393, 187-192. doi: 10.22059/ijvm.2014.51890
Ahani Azari, Ania, Zahraei Salehi, Taghi, Nayeri Fasaei, Bahar, Madadgar, Omid, Alebouyeh, Masoud. (1393). 'Construction of a recombinant vector for site-directed mutagenesis in Salmonella typhimurium', , 8(3), pp. 187-192. doi: 10.22059/ijvm.2014.51890
Ahani Azari, Ania, Zahraei Salehi, Taghi, Nayeri Fasaei, Bahar, Madadgar, Omid, Alebouyeh, Masoud. Construction of a recombinant vector for site-directed mutagenesis in Salmonella typhimurium. , 1393; 8(3): 187-192. doi: 10.22059/ijvm.2014.51890

Construction of a recombinant vector for site-directed mutagenesis in Salmonella typhimurium

Iranian Journal of Veterinary Medicine
مقاله 5، دوره 8، شماره 3، دی 2014، صفحه 187-192    اصل مقاله (183.88 K)
شناسه دیجیتال (DOI): 10.22059/ijvm.2014.51890
نویسندگان
Ania Ahani Azari1؛ Taghi Zahraei Salehi* 1؛ Bahar Nayeri Fasaei1؛ Omid Madadgar1؛ Masoud Alebouyeh2
1Department of Microbiology, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran
2Bacis and Molecular Epidemiology of Gastrointestinal Disorders Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran
چکیده
BACKGROUND: Among all common techniques in sitedirected
mutagenesis, λ Red recombinase system has been
widely used to knock out chromosomal genes in bacteria. In this
method, there is always the risk of DNA Linear digestion by
host's restriction enzymes that leads to the low frequency of
recombination. OBJECTIVES:To overcome this, we constructed
a recombinant vector to disrupt phoP gene in Salmonella
typhimurium. METHODS: The SOEing PCR method and
restriction enzymes were used to construct the vector. RESULTS:
The resulting plasmid, pTAAZ92, contains a Kanamycin
cassette with two long homologous arms flanking of the phoP
gene. CONCLUSIONS: After electrotransformation of the
pTAAZ92 into the Salmonella typhimurium , the phoP gene is
replaced by the Kanamycin cassette through homologous
recombination. According to the high homology of the phoP
gene in many of Salmonella species the pTAAZ92 can be used to
disrupt the phoP gene in most of these species.
کلیدواژه‌ها
gene disruption؛ Kanamycin cassette؛ Salmonella typhimurium؛ sitedirected mutagenesis
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