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R esearch
(Published online:
14-05-2015)
9.
Cloning and sequencing of hfq (host
factor required for synthesis of bacteriophage Q beta RNA) gene of
Salmonella Typhimurium isolated from poultry -
Parthasarathi Behera, Muhammed Kutty, Bhaskar Sharma, Ajay Kumar
and Meeta Saxena
Veterinary World, 8(5): 610-614
doi:
10.14202/vetworld.2015.610-614
Parthasarathi Behera:
Division of Biochemistry, Indian Veterinary Research Institute,
Izatnagar, Bareilly, Uttar Pradesh, India;
partha_vet@yahoo.co.in
Muhammed Kutty:
Division of Biochemistry, Indian Veterinary Research Institute,
Izatnagar, Bareilly, Uttar Pradesh, India;
mohd.kuttyvet@gmail.com
Bhaskar Sharma:
Division of Biochemistry, Indian Veterinary Research Institute,
Izatnagar, Bareilly, Uttar Pradesh, India; bhaskar@ivri.res.in
Ajay
Kumar:
Division of Biochemistry, Indian Veterinary Research Institute,
Izatnagar, Bareilly, Uttar Pradesh, India;
ajayivri@gmail.com
Meeta Saxena: Division of Biochemistry, Indian Veterinary
Research Institute, Izatnagar, Bareilly, Uttar Pradesh, India;
meeta@ivri.res.in
Received: 24-02-2015, Revised: 06-04-2015, Accepted: 15-04-2015,
Published online: 14-05-2015
Corresponding author:
Parthasarathi Behera, e-mail: partha_vet@yahoo.co.in
Citation:
Behera P, Kutty M,
Sharma B, Kumar A, Saxena M (2015) Cloning and sequencing of
hfq (host factor required for synthesis of bacteriophage Q
beta RNA) gene of Salmonella Typhimurium isolated from
poultry, Veterinary World, 8(5):610-614.
Abstract
Aim:
The aim was to clone and
sequence hfq
gene of
Salmonella Typhimurium strain
PM-45 and compare its sequence with hfq
gene of other serovar of
Salmonella.
Materials and Methods: Salmonella
Typhimurium strain PM-45 was procured
from the G. B. Pant University of Agriculture and Technology,
Pantnagar, India. The genomic DNA was isolated from
Salmonella
Typhimurium. Hfq
gene was polymerase chain reaction (PCR)
amplified from the DNA using specific primers, which was
subsequently cloned into pET32a vector and transformed into
Escherichia coli
BL21 pLys cells. The recombinant plasmid
was isolated and subjected to restriction enzyme digestion as well
as PCR. The clone was then sequenced. The sequence was analyzed
and submitted in GenBank.
Results: PCR produced an
amplicon of 309 bp. Restriction digestion of the recombinant
plasmid released the desired insert. The
hfq sequence shows 100%
homology with similar sequences from other
Salmonella Typhimurium
isolates. Both nucleotide and amino acid sequences are highly
conserved. The submitted sequence is having Genbank accession no
KM998764.
Conclusion: Hfq , the
hexameric RNA binding protein is one of the most important
post-transcriptional regulator of bacteria. The sequence of
hfq gene of
Salmonella
Typhimurium is highly conserved within
and between Salmonella enterica
serovars. This gene sequence is
probably under heavy selection pressure to maintain the
conformational integrity of its product in spite of its being not
a survival gene.
Keywords:
cloning, hfq,
RNA binding protein, sequencing,
Salmonella Typhimurium.
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