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.com
Volume 8
Journal of Biotechnology & Biomaterials
ISSN: 2155-952X
Biotech Congress 2018 & Enzymology 2018
March 05-07, 2018
JOINT EVENT
20
th
Global Congress on
Biotechnology
3
rd
International Conference on
Enzymology and Molecular Biology
&
March 05-07, 2018 London, UK
Mutated and wild type Gossypium universal stress
protein-2
(GUSP-2) gene confers resistance to
stresses in
Escherichia coli, Pichia pastoris
and cotton plant
Nadeem Hafeez
Center for Excellence in Molecular Biology, Pakistan
G
ossypium arboreum
is considered to be a rich source of stress responsive genes and EST data base revealed that most of
its genes are uncharacterized. The full length
Gossypium universal stress protein-2
(GUSP-2) gene (510bp) was cloned in
E. coli, Pichia pastoris and
Gossypium hirsutum
, characterized and point mutated at three positions separately at 352-354,
Lysine-60 to proline (
M1-usp-2
) and 214-216, aspartic acid-26 to serine (
M2-usp-2
) and 145-147, Lysine-3 to proline (
M3-
usp-2
) to study its role in abiotic stress tolerance. It was found that heterologous expression of one mutant (
M1-usp-2
) provided
enhanced tolerance against salt and osmotic stresses, recombinant cells have higher growth up to 10-5dilution in spot assay
as compared to
Wusp- 2
(wild type
GUSP-2
),
M2-usp-2
and
M3-usp-2
genes.
M1-usp-2
in Pichia pastoris transcript profiling
exhibited significant expression (7.1-fold) to salt and (9.7) and osmotic stresses.
M1-usp-2
gene was also found to enhance
drought tolerance and significant expression (8.7) in CIM-496-
Gossypium hirsutum
transgenic plants. However, little tolerance
against heat and cold stresses both in recombinant yeast and bacterial cells was observed. The results from our study concluded
that activity of
GUSP-2
was enhanced in
M1-usp-2
but wipe out in
M2-usp-2
and
M3-usp-2
response remained almost parallel
to
W-usp-2
. Further, it was predicted through in silico analysis that
M1-usp-2
,
W-usp-2
and
M3-usp-2
may be directly involved
in stress tolerance or function as signaling molecule to activate the stress adaptive mechanism.
chnadeemhfz@gmail.comJ Biotechnol Biomater 2018, Volume 8
DOI: 10.4172/2155-952X-C2-092