<abstract xmlns="http://www.w3.org/1999/xhtml"><p>Bacteriophage GA coat protein formed self-assembly competent virus-like particles (VLPs) have been expressed previously in bacterial and yeast cells. On the basis of our previous experiments on the yeast vector pESC-URA / <italic>S. cerevisiae</italic> system containing two oppositely oriented promoters, new constructions were created with point-mutations in coat protein to mimic phage MS2-like RNA binding characteristics. Simultaneously, the MS2 operator sequence was added to mRNA desired for packaging. After the introduction of single-point mutations (S87N, K55N, R43K) and double-point mutations (S87N + K55N and S87N + R43K), the coat protein’s ability to form VLPs was retained, but yield from cells was decreased. Exchange of the 87<sup>th</sup> Ser to Asn in coat protein sequence in combination with bacteriophage MS2 translational operator provided specific packaging of the gene of interest (GFP). Although non-specific nucleic acid sequences were packaged, the remarkable specificity for packaging of the gene of interest can be achieved using the described approach.</p></abstract>

Bacteriophage GA coat protein formed self-assembly competent virus-like particles (VLPs) have been expressed previously in bacterial and yeast cells. On the basis of our previous experiments on the yeast vector pESC-URA / S. cerevisiae system containing two oppositely oriented promoters, new constructions were created with point-mutations in coat protein to mimic phage MS2-like RNA binding characteristics. Simultaneously, the MS2 operator sequence was added to mRNA desired for packaging. After the introduction of single-point mutations (S87N, K55N, R43K) and double-point mutations (S87N + K55N and S87N + R43K), the coat protein’s ability to form VLPs was retained, but yield from cells was decreased. Exchange of the 87th Ser to Asn in coat protein sequence in combination with bacteriophage MS2 translational operator provided specific packaging of the gene of interest (GFP). Although non-specific nucleic acid sequences were packaged, the remarkable specificity for packaging of the gene of interest can be achieved using the described approach.

<italic>In Vivo</italic> Packaging of mRNA in Yeast-Produced Bacteriophage GA Derived Virus-Like Particles

Latvian Biomedical Research and Study Centre

Proceedings of the Latvian Academy of Sciences. Section B. Natural, Exact, and Applied Sciences.

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

 Packaging of mRNA in Yeast-Produced Bacteriophage GA Derived Virus-Like Particles

Bacteriophage GA coat protein formed self-assembly competent virus-like particles (VLPs) have been expressed previously in bacterial and yeast cells. On the...

In Vivo Packaging of mRNA in Yeast-Produced Bacteriophage GA Derived Virus-Like Particles

Online veröffentlicht: 09. Sept. 2017

Seitenbereich: 266 - 274

Eingereicht: 11. Juni 2015

Akzeptiert: 26. Mai 2017

DOI: https://doi.org/10.1515/prolas-2017-0045

Schlüsselwörter
virus-like particles, GA, MS2, operator

© 2017 Dagnija Ārgule et al., published by De Gruyter Open

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

In Vivo Packaging of mRNA in Yeast-Produced Bacteriophage GA Derived Virus-Like Particles

Online veröffentlicht: 09. Sept. 2017

Seitenbereich: 266 - 274

Eingereicht: 11. Juni 2015

Akzeptiert: 26. Mai 2017

DOI: https://doi.org/10.1515/prolas-2017-0045

Schlüsselwörtervirus-like particles, GA, MS2, operator

© 2017 Dagnija Ārgule et al., published by De Gruyter Open

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Schlüsselwörter
virus-like particles, GA, MS2, operator