Uticaj marker gena za kanamicinsku rezistenciju na stabilnost 2μ ekspresionih plazmida
Effect of the kanamycin resistance marker on stability of 2μ-based expression plasmids
Abstract
U ovom radu opisan je uticaj gena za kanamicinsku rezistenciju (Kmr) na održavanje 2μm plazmida u Saccharomyces cerevisiae. Prisustvo ovog marker gena dovodi do gubitka stabilnog model-vektora konstantnom stopom nezavisnom od izvora ugljenika i stope rasta kulture. Kod sojeva za sintezu heterolognih proteina sa galaktoznog promotora (GALUAS) uvođenje Kmr rezultira "čišćenjem" od kvaščevih epizomalnih plazmida (YEp) u svega nekoliko generacija. Primena selektivnog pritiska na sojeve koji proizvode rekombinantnu penicilin G amidazu (rPGA) nije dovela do očekivanog povećanja prinosa proteina. Ispitivanjem uticaja samih genetičkih elemenata za proizvodnju heterolognih proteina na stabilnost vektora pokazano je da najjače destabilišuće dejstvo ima prisustvo i eksprecija stranog gena.
In this paper we describe the effect of the kanamycin resistance gene (Kmr) on 2μm-based plasmid maintenance in Saccharomyces cerevisiae. The influence of this marker gene on the loss of the stable model-vectors proved to be constant, as well as independent of carbon source and culture growth rates. In strains for GALUAS - driven heterologous protein production introduction of Kmr resulted in curing of the yeast episomal plasmid (YEp) from the population in a small number of generations. Application of selective pressure on the strain producing recombinant penicillin G amidase (rPGA) did not provide the expected increase of protein yield. The influence of genetic elements for heterologous protein production on vector stability was examined, and the most destabilizing factors prove to be the presence and expression of the foreign gene.
Keywords:
selective markers / Saccharomyces cerevisiae / plasmid stability / penicillin G amidase / kanamycin resistance / 2μm plasmidSource:
Archives of Biological Sciences, 2007, 59, 1, 1-12Publisher:
- Srpsko biološko društvo, Beograd, i dr.
Collections
Institution/Community
Institut za molekularnu genetiku i genetičko inženjerstvoTY - JOUR AU - Stanković, Nada AU - Vasiljević, Branka AU - Ljubijankić, Goran PY - 2007 UR - https://imagine.imgge.bg.ac.rs/handle/123456789/282 AB - U ovom radu opisan je uticaj gena za kanamicinsku rezistenciju (Kmr) na održavanje 2μm plazmida u Saccharomyces cerevisiae. Prisustvo ovog marker gena dovodi do gubitka stabilnog model-vektora konstantnom stopom nezavisnom od izvora ugljenika i stope rasta kulture. Kod sojeva za sintezu heterolognih proteina sa galaktoznog promotora (GALUAS) uvođenje Kmr rezultira "čišćenjem" od kvaščevih epizomalnih plazmida (YEp) u svega nekoliko generacija. Primena selektivnog pritiska na sojeve koji proizvode rekombinantnu penicilin G amidazu (rPGA) nije dovela do očekivanog povećanja prinosa proteina. Ispitivanjem uticaja samih genetičkih elemenata za proizvodnju heterolognih proteina na stabilnost vektora pokazano je da najjače destabilišuće dejstvo ima prisustvo i eksprecija stranog gena. AB - In this paper we describe the effect of the kanamycin resistance gene (Kmr) on 2μm-based plasmid maintenance in Saccharomyces cerevisiae. The influence of this marker gene on the loss of the stable model-vectors proved to be constant, as well as independent of carbon source and culture growth rates. In strains for GALUAS - driven heterologous protein production introduction of Kmr resulted in curing of the yeast episomal plasmid (YEp) from the population in a small number of generations. Application of selective pressure on the strain producing recombinant penicillin G amidase (rPGA) did not provide the expected increase of protein yield. The influence of genetic elements for heterologous protein production on vector stability was examined, and the most destabilizing factors prove to be the presence and expression of the foreign gene. PB - Srpsko biološko društvo, Beograd, i dr. T2 - Archives of Biological Sciences T1 - Uticaj marker gena za kanamicinsku rezistenciju na stabilnost 2μ ekspresionih plazmida T1 - Effect of the kanamycin resistance marker on stability of 2μ-based expression plasmids EP - 12 IS - 1 SP - 1 VL - 59 DO - 10.2298/ABS0701001S ER -
@article{ author = "Stanković, Nada and Vasiljević, Branka and Ljubijankić, Goran", year = "2007", abstract = "U ovom radu opisan je uticaj gena za kanamicinsku rezistenciju (Kmr) na održavanje 2μm plazmida u Saccharomyces cerevisiae. Prisustvo ovog marker gena dovodi do gubitka stabilnog model-vektora konstantnom stopom nezavisnom od izvora ugljenika i stope rasta kulture. Kod sojeva za sintezu heterolognih proteina sa galaktoznog promotora (GALUAS) uvođenje Kmr rezultira "čišćenjem" od kvaščevih epizomalnih plazmida (YEp) u svega nekoliko generacija. Primena selektivnog pritiska na sojeve koji proizvode rekombinantnu penicilin G amidazu (rPGA) nije dovela do očekivanog povećanja prinosa proteina. Ispitivanjem uticaja samih genetičkih elemenata za proizvodnju heterolognih proteina na stabilnost vektora pokazano je da najjače destabilišuće dejstvo ima prisustvo i eksprecija stranog gena., In this paper we describe the effect of the kanamycin resistance gene (Kmr) on 2μm-based plasmid maintenance in Saccharomyces cerevisiae. The influence of this marker gene on the loss of the stable model-vectors proved to be constant, as well as independent of carbon source and culture growth rates. In strains for GALUAS - driven heterologous protein production introduction of Kmr resulted in curing of the yeast episomal plasmid (YEp) from the population in a small number of generations. Application of selective pressure on the strain producing recombinant penicillin G amidase (rPGA) did not provide the expected increase of protein yield. The influence of genetic elements for heterologous protein production on vector stability was examined, and the most destabilizing factors prove to be the presence and expression of the foreign gene.", publisher = "Srpsko biološko društvo, Beograd, i dr.", journal = "Archives of Biological Sciences", title = "Uticaj marker gena za kanamicinsku rezistenciju na stabilnost 2μ ekspresionih plazmida, Effect of the kanamycin resistance marker on stability of 2μ-based expression plasmids", pages = "12-1", number = "1", volume = "59", doi = "10.2298/ABS0701001S" }
Stanković, N., Vasiljević, B.,& Ljubijankić, G.. (2007). Uticaj marker gena za kanamicinsku rezistenciju na stabilnost 2μ ekspresionih plazmida. in Archives of Biological Sciences Srpsko biološko društvo, Beograd, i dr.., 59(1), 1-12. https://doi.org/10.2298/ABS0701001S
Stanković N, Vasiljević B, Ljubijankić G. Uticaj marker gena za kanamicinsku rezistenciju na stabilnost 2μ ekspresionih plazmida. in Archives of Biological Sciences. 2007;59(1):1-12. doi:10.2298/ABS0701001S .
Stanković, Nada, Vasiljević, Branka, Ljubijankić, Goran, "Uticaj marker gena za kanamicinsku rezistenciju na stabilnost 2μ ekspresionih plazmida" in Archives of Biological Sciences, 59, no. 1 (2007):1-12, https://doi.org/10.2298/ABS0701001S . .