A second generation snp-derived Escherichia coli-Streptomyces shuttle expression vector that is generally transferable by conjugation
Abstract
An Escherichia coli-Streptomyces shuttle vector (pJN100) was constructed, by inserting an origin of transfer (oriT), derived from the E coli broad host range plasmid RK2, into pANT1202, a high-copy-number vector for gene expression in Streptomyces. The resulting conjugably transferable vector contains the pANT1202-derived SnpR (LysR-like protein) activated snpA promoter that drives strong heterologous expression of proteins. We initially demonstrated that plasmid pJN 100 was transferred with high frequency (10(-5-7) exconjugants per recipient) into several Streptomyces strains that were refractory to transformation by other means. Plasmid pJN100 was also shown to be stable in E. coli and Streptomyces. We confirmed functional protein expression by using a pJN100 derivative to complement a mutant of Streptomyces griseus with a disrupted chromosomal copy of the gene nonM, a gene encoding an essential reductase in the nonactin biosynthesis gene cluster. High levels of protein expression we...re confirmed using Western blotting to assess the production of the serine esterase NonR, an enzyme responsible for nonactin resistance in the nonactin producer S. griseus. (c) 2006 Elsevier Inc. All rights reserved.
Keywords:
Streptomyces / conjugation / shuttle vector / oriT / snpA promoter / RK2 / heterologous expressionSource:
Plasmid, 2006, 56, 3, 223-227Publisher:
- Academic Press Inc Elsevier Science, San Diego
DOI: 10.1016/j.plasmid.2006.05.002
ISSN: 0147-619X
PubMed: 16806469
WoS: 000242306000008
Scopus: 2-s2.0-33750453762
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Inovacioni centar / Innovation CentreTY - JOUR AU - Nikodinović-Runić, Jasmina AU - Priestley, Nigel D. PY - 2006 UR - https://cherry.chem.bg.ac.rs/handle/123456789/764 AB - An Escherichia coli-Streptomyces shuttle vector (pJN100) was constructed, by inserting an origin of transfer (oriT), derived from the E coli broad host range plasmid RK2, into pANT1202, a high-copy-number vector for gene expression in Streptomyces. The resulting conjugably transferable vector contains the pANT1202-derived SnpR (LysR-like protein) activated snpA promoter that drives strong heterologous expression of proteins. We initially demonstrated that plasmid pJN 100 was transferred with high frequency (10(-5-7) exconjugants per recipient) into several Streptomyces strains that were refractory to transformation by other means. Plasmid pJN100 was also shown to be stable in E. coli and Streptomyces. We confirmed functional protein expression by using a pJN100 derivative to complement a mutant of Streptomyces griseus with a disrupted chromosomal copy of the gene nonM, a gene encoding an essential reductase in the nonactin biosynthesis gene cluster. High levels of protein expression were confirmed using Western blotting to assess the production of the serine esterase NonR, an enzyme responsible for nonactin resistance in the nonactin producer S. griseus. (c) 2006 Elsevier Inc. All rights reserved. PB - Academic Press Inc Elsevier Science, San Diego T2 - Plasmid T1 - A second generation snp-derived Escherichia coli-Streptomyces shuttle expression vector that is generally transferable by conjugation VL - 56 IS - 3 SP - 223 EP - 227 DO - 10.1016/j.plasmid.2006.05.002 ER -
@article{ author = "Nikodinović-Runić, Jasmina and Priestley, Nigel D.", year = "2006", abstract = "An Escherichia coli-Streptomyces shuttle vector (pJN100) was constructed, by inserting an origin of transfer (oriT), derived from the E coli broad host range plasmid RK2, into pANT1202, a high-copy-number vector for gene expression in Streptomyces. The resulting conjugably transferable vector contains the pANT1202-derived SnpR (LysR-like protein) activated snpA promoter that drives strong heterologous expression of proteins. We initially demonstrated that plasmid pJN 100 was transferred with high frequency (10(-5-7) exconjugants per recipient) into several Streptomyces strains that were refractory to transformation by other means. Plasmid pJN100 was also shown to be stable in E. coli and Streptomyces. We confirmed functional protein expression by using a pJN100 derivative to complement a mutant of Streptomyces griseus with a disrupted chromosomal copy of the gene nonM, a gene encoding an essential reductase in the nonactin biosynthesis gene cluster. High levels of protein expression were confirmed using Western blotting to assess the production of the serine esterase NonR, an enzyme responsible for nonactin resistance in the nonactin producer S. griseus. (c) 2006 Elsevier Inc. All rights reserved.", publisher = "Academic Press Inc Elsevier Science, San Diego", journal = "Plasmid", title = "A second generation snp-derived Escherichia coli-Streptomyces shuttle expression vector that is generally transferable by conjugation", volume = "56", number = "3", pages = "223-227", doi = "10.1016/j.plasmid.2006.05.002" }
Nikodinović-Runić, J.,& Priestley, N. D.. (2006). A second generation snp-derived Escherichia coli-Streptomyces shuttle expression vector that is generally transferable by conjugation. in Plasmid Academic Press Inc Elsevier Science, San Diego., 56(3), 223-227. https://doi.org/10.1016/j.plasmid.2006.05.002
Nikodinović-Runić J, Priestley ND. A second generation snp-derived Escherichia coli-Streptomyces shuttle expression vector that is generally transferable by conjugation. in Plasmid. 2006;56(3):223-227. doi:10.1016/j.plasmid.2006.05.002 .
Nikodinović-Runić, Jasmina, Priestley, Nigel D., "A second generation snp-derived Escherichia coli-Streptomyces shuttle expression vector that is generally transferable by conjugation" in Plasmid, 56, no. 3 (2006):223-227, https://doi.org/10.1016/j.plasmid.2006.05.002 . .