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3 Biotech

, 9:232 | Cite as

Development of a novel DsRed-NLS vector with a monopartite classical nuclear localization signal

  • Hee Sang You
  • Yeon Jeong Ok
  • Eun Jeong Lee
  • Sang Sun Kang
  • Sung Hee HyunEmail author
Original Article
  • 29 Downloads

Abstract

The nuclear localization signal (NLS) marks proteins for transport to the nucleus and is used in various applications in many fields. NLSs are used to achieve efficient and stable transport of biomolecules. Previously, commercial vectors used in NLS studies contained three iterations of the NLS sequence, but these sequences can affect experimental results and alter protein function. Here, we investigated a new vector using a single classical NLS sequence with a mutation in pDsRed2-C1-wt to reduce experimental artifacts. In the newly constructed pDsRed2-C1-1NLS vector, the NLS sequence is placed near the multiple cloning sites of pDsRed2-C1-wt, and the multiple cloning site region was designed to facilitate insertion of the desired gene by site-directed mutagenesis. Fluorescent protein expression in the nucleus can be visually confirmed. The results show that the fluorescent protein was bound to the transport protein. The constructed vector had a cell survival rate of 89–95% and a transfection efficiency of 39–56% when introduced into animal cells, which are similar to those of other NLS vectors. Additionally, the constructed NLS vector can be used to demonstrate complementary binding between target proteins, and that the target protein is transported by the NLS transport system. Especially, we show that the vector can be useful for experiments involving the S100A10 gene. In addition, the constructed vector is useful for studies of genes and proteins that show potential for gene therapy or drug delivery applications.

Keywords

Nuclear localization signal Vector Single construction of sequence Viability Transfection efficiency S100A10 

Notes

Acknowledgements

This research was supported by the Basic Science Research Program, through the National Research Foundation of Korea (NRF), funded by the Ministry of Science ICT & Future Planning (NRF-2014R1A1A1002349). This article was revised by a professional English proofreader (Editage).

Author contributions

HSY, YJO, EJL and SSK performed the conceptualization; HSY, YJO and EJL performed the data curation and formal analysis; SSK and SHH performed funding and acquisition and resources; HSY, YJO, EJL and SSK performed the experiments including of investigation, methodology, software; HSY wrote the manuscript including of data visualization, original draft, review and editing; SHH performed project administration, supervision and validation. All authors read and approved the final manuscript.

Funding

This research was supported by the Basic Science Research Program, through the National Research Foundation of Korea (NRF), funded by the Ministry of Science ICT & Future Planning (NRF-2014R1A1A1002349).

Compliance with ethical standards

Conflict of interest

The authors have no conflict of interest (commercial or otherwise) to declare regarding this study.

Supplementary material

13205_2019_1770_MOESM1_ESM.docx (2.6 mb)
Supplementary file1 (DOCX 2698 kb)

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Copyright information

© King Abdulaziz City for Science and Technology 2019

Authors and Affiliations

  • Hee Sang You
    • 1
    • 2
  • Yeon Jeong Ok
    • 2
  • Eun Jeong Lee
    • 3
  • Sang Sun Kang
    • 3
  • Sung Hee Hyun
    • 1
    • 2
    Email author
  1. 1.Department of Senior Healthcare, BK21 Plus Program, Graduate SchoolEulji UniversityDaejeonRepublic of Korea
  2. 2.Department of Biomedical Laboratory Science, School of MedicineEulji UniversityDaejeonRepublic of Korea
  3. 3.Department of Biology EducationChungbuk National UniversityCheongjuRepublic of Korea

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