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 date: March 2020Source: Materials Science and Engineering: C, Volume 108Author(s): Piotr M. Skowron, Natalia Krawczun, Joanna Zebrowska, Daria Krefft, Olga Zołnierkiewicz, Marta Bielawa, Joanna Jezewska-Frackowiak, Lukasz Janus, Malgorzata Witkowska, Malgorzata Palczewska, Adriana Schumacher, Anna Wardowska, Milena Deptula, Artur Czupryn, Piotr Mucha, Arkadiusz Piotrowski, Pawel Sachadyn, Sylwia Rodziewicz-Motowidlo, Michal Pikula, Agnieszka Zylicz-StachulaAbstractA 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 sequ...
Source: Materials Science and Engineering: C - Category: Materials Science Source Type: research