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Design of a synthetic miniR1 plasmid and its production by engineered Escherichia coli.
Bioprocess Biosyst Eng 2019; 42(8):1391-1397BB

Abstract

A synthetic plasmid consisting of the minimal elements for replication control of the R1 replicon and kanamycin resistance marker, which was named pminiR1, was developed. pminiR1 production was tested at 30 °C under aerobic and microaerobic conditions in Escherichia coli W3110 recA- (W1). The plasmid DNA yields from biomass (YpDNA/X) were only 0.06 ± 0.02 and 0.22 ± 0.11 mg/g under aerobic and microaerobic conditions, respectively. As an option to increase YpDNA/X values, pminiR1 was introduced in an engineered E. coli strain expressing the Vitreoscilla hemoglobin inserted in chromosome (W12). The YpDNA/X values using strain W12 increased to 0.85 ± 0.05 and 1.53 ± 0.14 mg/g under aerobic and microaerobic conditions, respectively. pminiR1 production in both strains was compared with that of pUC57Kan at 37 °C under aerobic and microaerobic conditions. The YpDNA/X values for pminiR1 using strain W12 were 6.25 ± 0.16 and 9.27 ± 0.95 mg/g under aerobic and microaerobic conditions, respectively. Such yields were similar to those obtained for plasmid pUC57Kan using strain W12 (6.9 ± 0.64 and 10.85 ± 1.06 mg/g for aerobic and microaerobic cultures, respectively). Therefore, the synthetic minimal plasmid based on the R1 replicon is a valuable alternative to pUC plasmids for biotechnological applications.

Authors+Show Affiliations

Departamento de Procesos y Tecnología, Universidad Autónoma Metropolitana-Cuajimalpa, Vasco de Quiroga 4871, Santa Fe, 05348, Mexico City, Mexico. alara@correo.cua.uam.mx.Posgrado en Ciencias Naturales e Ingeniería, Universidad Autónoma Metropolitana-Cuajimalpa, Vasco de Quiroga 4871, Santa Fe, 05348, Mexico City, Mexico.Posgrado en Ciencias Naturales e Ingeniería, Universidad Autónoma Metropolitana-Cuajimalpa, Vasco de Quiroga 4871, Santa Fe, 05348, Mexico City, Mexico.Posgrado en Ciencias Naturales e Ingeniería, Universidad Autónoma Metropolitana-Cuajimalpa, Vasco de Quiroga 4871, Santa Fe, 05348, Mexico City, Mexico.Departamento de Procesos y Tecnología, Universidad Autónoma Metropolitana-Cuajimalpa, Vasco de Quiroga 4871, Santa Fe, 05348, Mexico City, Mexico.Departamento de Procesos y Tecnología, Universidad Autónoma Metropolitana-Cuajimalpa, Vasco de Quiroga 4871, Santa Fe, 05348, Mexico City, Mexico.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

31006041

Citation

Lara, Alvaro R., et al. "Design of a Synthetic miniR1 Plasmid and Its Production By Engineered Escherichia Coli." Bioprocess and Biosystems Engineering, vol. 42, no. 8, 2019, pp. 1391-1397.
Lara AR, Velázquez D, Penella I, et al. Design of a synthetic miniR1 plasmid and its production by engineered Escherichia coli. Bioprocess Biosyst Eng. 2019;42(8):1391-1397.
Lara, A. R., Velázquez, D., Penella, I., Islas, F., González-De la Rosa, C. H., & Sigala, J. C. (2019). Design of a synthetic miniR1 plasmid and its production by engineered Escherichia coli. Bioprocess and Biosystems Engineering, 42(8), pp. 1391-1397. doi:10.1007/s00449-019-02129-2.
Lara AR, et al. Design of a Synthetic miniR1 Plasmid and Its Production By Engineered Escherichia Coli. Bioprocess Biosyst Eng. 2019;42(8):1391-1397. PubMed PMID: 31006041.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Design of a synthetic miniR1 plasmid and its production by engineered Escherichia coli. AU - Lara,Alvaro R, AU - Velázquez,Daniela, AU - Penella,Inés, AU - Islas,Fabiola, AU - González-De la Rosa,Claudia H, AU - Sigala,Juan-Carlos, Y1 - 2019/04/20/ PY - 2019/01/14/received PY - 2019/04/08/accepted PY - 2019/4/22/pubmed PY - 2019/4/22/medline PY - 2019/4/22/entrez KW - Microaerobic KW - Minimal plasmid KW - R1 replicon KW - Synthetic plasmid KW - Vitreoscilla hemoglobin SP - 1391 EP - 1397 JF - Bioprocess and biosystems engineering JO - Bioprocess Biosyst Eng VL - 42 IS - 8 N2 - A synthetic plasmid consisting of the minimal elements for replication control of the R1 replicon and kanamycin resistance marker, which was named pminiR1, was developed. pminiR1 production was tested at 30 °C under aerobic and microaerobic conditions in Escherichia coli W3110 recA- (W1). The plasmid DNA yields from biomass (YpDNA/X) were only 0.06 ± 0.02 and 0.22 ± 0.11 mg/g under aerobic and microaerobic conditions, respectively. As an option to increase YpDNA/X values, pminiR1 was introduced in an engineered E. coli strain expressing the Vitreoscilla hemoglobin inserted in chromosome (W12). The YpDNA/X values using strain W12 increased to 0.85 ± 0.05 and 1.53 ± 0.14 mg/g under aerobic and microaerobic conditions, respectively. pminiR1 production in both strains was compared with that of pUC57Kan at 37 °C under aerobic and microaerobic conditions. The YpDNA/X values for pminiR1 using strain W12 were 6.25 ± 0.16 and 9.27 ± 0.95 mg/g under aerobic and microaerobic conditions, respectively. Such yields were similar to those obtained for plasmid pUC57Kan using strain W12 (6.9 ± 0.64 and 10.85 ± 1.06 mg/g for aerobic and microaerobic cultures, respectively). Therefore, the synthetic minimal plasmid based on the R1 replicon is a valuable alternative to pUC plasmids for biotechnological applications. SN - 1615-7605 UR - https://www.unboundmedicine.com/medline/citation/31006041/Design_of_a_synthetic_miniR1_plasmid_and_its_production_by_engineered_Escherichia_coli_ L2 - https://dx.doi.org/10.1007/s00449-019-02129-2 DB - PRIME DP - Unbound Medicine ER -