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Genetic transformation of Chlorella vulgaris mediated by HIV-TAT peptide

Gadamchetty, Pavan, Mullapudi, Phanindra Lakshmi Venkata, Sanagala, Raghavendrarao, Markandan, Manickavasagam, Polumetla, Ananda Kumar
3 Biotech 2019 v.9 no.4 pp. 139
Chlorella vulgaris, fluorescence, fluorescence microscopes, genes, genetic transformation, green fluorescent protein, microalgae, octoxynol, peptides, polymerase chain reaction, transfer DNA, transferases
Scientific interest in microalgal species is growing and, genetic transformation has definitely opened more avenues, in the ongoing research on microphytes. In the present study, we have attempted to transform Chlorella vulgaris by mobilizing double-stranded linear Transfer DNA (T-DNA) comprised of green fluorescent protein (egfp) gene cassette and hygromycin phosphotransferase II (hptII) gene cassette non-covalently bound to TAT peptide, into C. vulgaris cells treated with Triton X-100. The transformed C. vulgaris cells when examined under fluorescent microscope, exhibited green fluorescence in comparison to the untransformed cells. The transformed cells were further screened, and the surviving colonies were sub-cultured, on BG11 medium fortified with Hygromycin. The surviving colonies were confirmed for the presence of integrated T-DNA by Polymerase Chain Reaction with egfp and hptII gene-specific primers. This methodology has potential to substitute the existing tedious transformation methodologies and ease the future studies in microalgae.