STANDARDIZATION OF REGENERATION AND TRANSFORMATION OF Coleus forskohlii WITH THE TRAIL GENE
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Date
2019-08-22
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UNIVERSITY OF AGRICULTURAL SCIENCES GKVK, BENGALURU
Abstract
Coleus forskohlii is a medicinal plant of perennial and aromatic nature. Cancer is
one of the leading causes of morbidity and mortality worldwide and the presently
available treatments for cancer have several serious side effects and costly. Bio pharming
has a great role in producing drugs at low cost. In the present investigation TNF Related
Apoptosis Inducing Ligand (TRAIL) gene was made to expressed in the Coleus forskohlii.
The standardization of regeneration protocol of Coleus forskohlii using leaf explants was
conducted. Various concentrations of growth hormones such as auxin (IAA) and
cytokinin (BAP) were tired to induce regeneration. Significantly highest percent of
callus, mean number of shoots per callus and the rooting was obtained in the combination
containing 9 mg L-1 of BAP and 0.4 mg L-1 of IAA. Screening of plant putative
transformants was done using PCR for gene integration and for the expression studies,
c-DNA analysis, direct plate ELISA, indirect dot immuonobinding assay (DIBA) and
SDS-PAGE analysis was carried out. PCR amplification showed the presence of 400bp
of TRAIL gene amplicon in transformed plants and the protein size obtained through
SDS-PAGE analysis was about 30kDa.