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Research
(Published
online: 25-09-2014)
17.
Cloning and sequencing of protein L-isoaspartyl O-methyl
transferase of Salmonella Typhimurium isolated from poultry
-
S. K. Dixit, D. P.
Hota, M. Kumawat, T. K. Goswami and M. Mahawar
Veterinary World, 7(9): 712-716
doi:
10.14202/vetworld.2014.712-716
S. K.
Dixit:
Immunology Section, Indian Veterinary Research Institute,
Izatnagar, Bareilly, Uttar Pradesh, India;
sunildixit1987@gmail.com
D. P.
Hota:
Division of Biochemistry, Indian Veterinary Research Institute,
Izatnagar, Bareilly, Uttar Pradesh, India;
durgaprasad.hota04@gmail.com
M.
Kumawat:
Division of Biochemistry, Indian Veterinary Research Institute,
Izatnagar, Bareilly, Uttar Pradesh, India;
0711mworld@gmail.com
T. K.
Goswami:
Immunology Section, Indian Veterinary Research Institute,
Izatnagar, Bareilly, Uttar Pradesh, India;
goswami.tapas@gmail.com
M.
Mahawar:
Division of Biochemistry, Indian Veterinary Research Institute,
Izatnagar, Bareilly, Uttar Pradesh, India;
manishbiochemistry@gmail.com
Received:
16-05-2014, Revised:
28-07-2014, Accepted:
04-08-2014, Published online:
25-09-2014
Corresponding author:
S. K. Dixit, e-mail: sunildixit1987@gmail.com
Abstract
Aim:
To clone the Salmonella Typhimurium protein L-isoaspartyl
O-methyl transferase (PIMT) enzyme and to analyze
the sequence with PIMT gene of other pathogenic serovars of
Salmonella.
Materials and Methods: Salmonella Typhimurium strain
E-2375 was procured from the National Salmonella Center, IVRI. The
genomic DNA was isolated from Salmonella Typhimurium.
Polymerase chain reaction (PCR) was carried out to amplify PIMT
gene using the designed primers. The PCR product was cloned
into pET28c plasmid vector and transformed into Escherichia
coli DH5α cells. The plasmid was isolated from E. coli
and was sequenced. The sequence was analyzed and submitted in
Genbank.
Results: The PCR product revealed a distinct amplicon of 627
bp. The clone was confirmed by PCR. Sequencing data revealed 100%
homology between PIMT sequences from Salmonella
Typhimurium strain E-2375 (used in the current study) and
PIMT sequences of standard reported strain (Salmonella
Typhimurium str. LT2) in NCBI data base. This submitted
sequence in Genbank having accession no. KJ575536.
Conclusions: PIMT gene of Salmonella is highly
conserved in most of the pathogenic Salmonella serovars.
The PIMT clone can be used to isolate PIMT protein.
This PIMT protein will be helpful to identify isoaspartate
containing proteins thus can help in study Salmonella
virulence.
Keywords: cloning, sequencing, Salmonella
Typhimurium protein L-isoaspartyl O-methyl
transferase, virulence.
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