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Volume 7, Issue 4 (Suppl)
J Biotechnol Biomater, an open access journal
ISSN: 2155-952X
Bio America 2017
October 19-20, 2017
October 19-20, 2017 | New York, USA
18
th
Biotechnology Congress
Mutated and wild type Gossypium universal stress protein-2 (GUSP-2) gene confers resistance to abiotic
stresses in transgenic cotton plant
Nadeem Hafeez
Center for Excellence in Molecular Biology (CEMB), Pakistan
G
ossypium arboreum
is considered to be a rich source of stress responsive genes and EST data base revealed that mostly 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)
& 145-147, Lysine-3 to proline
(M3-usp-2)
to study its role
in abiotic stress tolerance. It was found that heterologus 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
W-usp-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 2017, 7:4 (Suppl)
DOI: 10.4172/2155-952X-C1-080