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3 Biotech

, 9:139 | Cite as

Genetic transformation of Chlorella vulgaris mediated by HIV-TAT peptide

  • Pavan GadamchettyEmail author
  • Phanindra Lakshmi Venkata Mullapudi
  • Raghavendrarao Sanagala
  • Manickavasagam Markandan
  • Ananda Kumar Polumetla
Original Article
  • 71 Downloads

Abstract

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.

Keywords

Chlorella vulgaris HIV-TAT Cell-penetrating peptide Genetic transformation Microalgae 

Notes

Acknowledgements

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.

Author contributions

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.

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Copyright information

© King Abdulaziz City for Science and Technology 2019

Authors and Affiliations

  1. 1.National Research Centre on Plant BiotechnologyNew DelhiIndia
  2. 2.Department of Biotechnology and Genetic Engineering, School of BiotechnologyBharathidasan UniversityTiruchirappalliIndia
  3. 3.Visargha Agri Sciences Private LimitedBhubaneswarIndia
  4. 4.Ganga Kaveri Seeds Private LimitedHyderabadIndia
  5. 5.Indian Institute of Rice ResearchHyderabadIndia

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