A vector-enzymatic DNA fragment amplification-expression technology for construction of artificial, concatemeric DNA, RNA and proteins for novel biomaterials, biomedical and industrial applications - Publication - MOST Wiedzy

Search

A vector-enzymatic DNA fragment amplification-expression technology for construction of artificial, concatemeric DNA, RNA and proteins for novel biomaterials, biomedical and industrial applications

Abstract

A DNA fragment amplification/expression technology for the production of new generation biomaterials for scientific, industrial and biomedical applications is described. The technology enables the formation of artificial Open Reading Frames (ORFs) encoding concatemeric RNAs and proteins. It recruits the Type IIS SapI restriction endonuclease (REase) for an assembling of DNA fragments in an ordered head-to-tail-orientation. The technology employs a vector-enzymatic system, dedicated to the expression of newly formed, concatemeric ORFs from strong promoters. Four vector series were constructed to suit specialised needs. As a proof of concept, a model amplification of a 7-amino acid (aa) epitope from the S protein of HBV virus was performed, resulting in 500 copies of the epitope-coding DNA segment, consecutively linked and expressed in Escherichia coli (E. coli). Furthermore, a peptide with potential pro-regenerative properties (derived from an angiopoietin-related growth factor) was designed. Its aa sequence was back-translated, codon usage optimized and synthesized as a continuous ORF 10-mer. The 10-mer was cloned into the amplification vector, enabling the N-terminal fusion and multiplication of the encoded protein with MalE signal sequence. The obtained genes were expressed, and the proteins were purified. Conclusively, we show that the proteins are neither cytotoxic nor immunogenic and they have a very low allergic potential.

Citations

  • 3

    CrossRef

  • 2

    Web of Science

  • 2

    Scopus

Authors (20)

  • Photo of prof. dr hab. Piotr Skowron

    Piotr Skowron prof. dr hab.

  • Photo of  Natalia Krawczun

    Natalia Krawczun

  • Photo of  Joanna Żebrowska

    Joanna Żebrowska

  • Photo of  Daria Krefft

    Daria Krefft

  • Photo of  Olga Żołnierkiewicz

    Olga Żołnierkiewicz

  • Photo of  Marta Bielawa

    Marta Bielawa

  • Photo of  Joanna Jeżewska-Frąckowiak

    Joanna Jeżewska-Frąckowiak

  • Photo of dr Łukasz Janus

    Łukasz Janus dr

  • Photo of  Małgorzata Witkowska

    Małgorzata Witkowska

  • Photo of  Małgorzata Palczewska

    Małgorzata Palczewska

  • Photo of mgr Adriana Schumacher

    Adriana Schumacher mgr

  • Photo of  Anna Wardowska

    Anna Wardowska

  • Photo of  Milena Deptuła

    Milena Deptuła

  • Photo of dr Artur Czupryn

    Artur Czupryn dr

  • Photo of dr hab. Piotr Mucha

    Piotr Mucha dr hab.

  • Photo of dr hab. Arkadiusz Piotrowski

    Arkadiusz Piotrowski dr hab.

  • Photo of dr hab. Sylwia Rodziewicz-Motowidło

    Sylwia Rodziewicz-Motowidło dr hab.

  • Photo of  Michal Pikuła

    Michal Pikuła

  • Photo of  Agnieszka Zylicz-Stachula

    Agnieszka Zylicz-Stachula

Details

Category:
Articles
Type:
artykuły w czasopismach
Published in:
Materials Science & Engineering C-Materials for Biological Applications no. 108,
ISSN: 0928-4931
Language:
English
Publication year:
2020
Bibliographic description:
Skowron P., Krawczun N., Żebrowska J., Krefft D., Żołnierkiewicz O., Bielawa M., Jeżewska-Frąckowiak J., Janus Ł., Witkowska M., Palczewska M., Schumacher A., Wardowska A., Deptuła M., Czupryn A., Mucha P., Piotrowski A., Sachadyn P., Rodziewicz-Motowidło S., Pikuła M., Zylicz-Stachula A.: A vector-enzymatic DNA fragment amplification-expression technology for construction of artificial, concatemeric DNA, RNA and proteins for novel biomaterials, biomedical and industrial applications// Materials Science & Engineering C-Materials for Biological Applications -Vol. 108, (2020), s.110426-
DOI:
Digital Object Identifier (open in new tab) 10.1016/j.msec.2019.110426
Verified by:
Gdańsk University of Technology

seen 31 times

Recommended for you

Meta Tags