Isolation
of plasmid from a Gram negative bacteria
Aeromonas
dhakensis strain F2S2-1 and its partial characterization
M.
Nadiga1, 2 V.V. Vaidyanathan1 and T. Thayumanavan2*
1Syngene
International Ltd Bangalore, India
2Department of
Biotechnology, Dr. G. R Damodaran College of Science, Civil Aerodrome Post,
Coimbatore – 641014, India
*Corresponding
Author E-mail: thayumanavan@yahoo.com
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Key
words
Aeromonas dhakensis
DNA nicking enzyme
Endonuclease
Plasmid sequencing
Publication Data
Paper
received : 30.06.2016
Revised
received : 12.08.2016
Re-revised
received : 20.01.2017
Accepted : 28.01.2017
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Abstract
Aim: A plasmid was
isolated from Aeromonas dhakensis strain F2S2-1 to understand their
function in the host. The main objective of the study was to isolate and
sequence the plasmid, predict genes and proteins encoded and to
experimentally prove their biochemical function.
Methodology:
The
Aeromonas dhakensis strain F2S2-1 plasmid was sequenced by Next
Generation Sequencing (NGS) on Illumina Miseq platform. Bioinformatics
analysis was carried out to predict the putative gene and encoded protein
functions. The NspV like endonuclease was recombinantly expressed in Escherichia
coli, partially purified and characterized for activity on plasmid
substrates.
Results:
Plasmid
sequencing and bioinformatics analysis predicted the presence of NspV like
endonuclease. Recombinant expression, purification and characterization of
NspV like endonuclease revealed the Nicking endonuclease activity of the
protein with plasmid substrates.
Interpretation:
The
NspV like endonuclease described in this study has DNA nicking activity. This
study would help in understanding better survival of Aeromonas sp. in
its ecosystem and the role of host plamids.
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conclusions enforced or derived, rest completely with the author(s).
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