Genetic transformation of Chlorella vulgaris mediated by HIV-TAT peptide
- 71 Downloads
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.
KeywordsChlorella vulgaris HIV-TAT Cell-penetrating peptide Genetic transformation Microalgae
The authors thankfully acknowledge the support from The Centre for Conservation and Utilization of Blue Green Algae (CCUBGA), Division of Microbiology, IARI, New Delhi and technical staff at NRCPB, IARI, New Delhi.
The experiments were conceived and designed by PAK, MLVP and GP. The experiments were performed by GP and SR. The data were analyzed by MM and SR. The reagents, materials and analysis tools were provided by PAK. The manuscript was prepared by GP and MLVP.
Compliance with ethical standards
Conflict of interest
The authors declare that there is no conflict of interest.
- Sambrook J, Russell DW (2001) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, New YorkGoogle Scholar
- Sambrook J, Fritschi EF, Maniatis T (1989) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, New YorkGoogle Scholar
- Ziemienowicz A, Tzfira T, Hohn B (2008) Mechanisms of T-DNA integration. Agrobacterium: from biology to biotechnology. Springer, New York, pp 396–441Google Scholar