Journal: Journal of Biological Inorganic Chemistry

Article Title: Distinctive structural properties of THB11, a pentacoordinate Chlamydomonas reinhardtii truncated hemoglobin with N- and C-terminal extensions

doi: 10.1007/s00775-020-01759-2

Figure Lengend Snippet: THB11 length variants form different oligomeric states. a Anion exchange chromatography (AEC) profiles of the indicated protein variants, which had been purified from E. coli cell lysates by Strep -tag affinity chromatography before. The brackets above the peaks indicate the elution fractions that were combined for subsequent analyses [black, blue: peak (P) 1, 2]. The molecular mass of each protein calculated from the primary sequence are indicated to the left. b Denaturing (SDS) and native gel electrophoreses of the protein solutions collected from the AEC peaks P1 and P2. The numbered bands observed in the native gels were cut out and subsequently subjected to denaturing gel electrophoresis. c Size exclusion chromatography (SEC) profiles of the proteins present in the two AEC peaks. kDa labels within the SEC graphs indicate the molecular masses calculated from the elution volumes according to the column’s calibration curve, colors indicate the original AEC P1 (black) or P2 (blue). d UV–Vis spectra of the protein solutions collected from the two AEC peaks, P1 and P2. The spectra were normalized with regard to the Soret maxima (set to 1)

Article Snippet: Cloning was done using E. coli strain DH5α MCR grown in LB Broth (Lennox) or on LB Agar (Lennox) from Carl Roth GmbH ( https://www.carlroth.com ) supplied with 100 μg × ml −1 ampicillin at 37 °C.

Techniques: Chromatography, Purification, Strep-tag, Affinity Chromatography, Sequencing, Nucleic Acid Electrophoresis, Size-exclusion Chromatography

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: ColE1-Plasmid Production in Escherichia coli : Mathematical Simulation and Experimental Validation

doi: 10.3389/fbioe.2015.00127

Figure Lengend Snippet: Growth curve for E. coli DH5α-pCMV-lacZ . The growth curve based on OD 600 measurements is marked by small black circles and the appropriate best-fit curve is indicated in blue. The harvesting time points T 1 , T 2 , and T 3 are presented as bold big points. The minimum–maximum area, within which the measurement values have to reside, is bordered by the red curve (minimal measured OD 600 value) and the green curve (maximal measured OD 600 value).

Article Snippet: The Escherichia coli strain DH5α (F–Φ80 lac ZΔM15 Δ( lac ZYA- arg F) U169 rec A1 end A1 hsd R17 (rK−, mK+) pho A sup E44 λ− thi -1 gyr A96 rel A1) (Source: Plasmid Factory, Bielefeld, Germany) was used as a host strain for transformation of the high copy plasmid pCMV-lacZ (Source: Plasmid Factory, Bielefeld, Germany) as well as the low copy plasmid pSUP 201-3 (Simon et al., ). pCMV-lacZ is a ColE1-derived high copy plasmid for therapeutic pDNA production with a size of 7164 bp.

Techniques:

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: ColE1-Plasmid Production in Escherichia coli : Mathematical Simulation and Experimental Validation

doi: 10.3389/fbioe.2015.00127

Figure Lengend Snippet: Growth curve for E. coli DH5α-pSUP 201-3 . The growth curve based on OD 600 measurements are marked by black circles and the appropriate best-fit curve is indicated in blue. The harvesting time points T 1 , T 2 , and T 3 are presented as bold big points. The minimum–maximum area, within which the measurement values have to reside, is bordered by the red curve (minimal measured OD 600 values) and the green curve (maximal measured OD 600 values).

Article Snippet: The Escherichia coli strain DH5α (F–Φ80 lac ZΔM15 Δ( lac ZYA- arg F) U169 rec A1 end A1 hsd R17 (rK−, mK+) pho A sup E44 λ− thi -1 gyr A96 rel A1) (Source: Plasmid Factory, Bielefeld, Germany) was used as a host strain for transformation of the high copy plasmid pCMV-lacZ (Source: Plasmid Factory, Bielefeld, Germany) as well as the low copy plasmid pSUP 201-3 (Simon et al., ). pCMV-lacZ is a ColE1-derived high copy plasmid for therapeutic pDNA production with a size of 7164 bp.

Techniques:

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: ColE1-Plasmid Production in Escherichia coli : Mathematical Simulation and Experimental Validation

doi: 10.3389/fbioe.2015.00127

Figure Lengend Snippet: Measured RNAI-, RNAII-, and plasmid concentrations for DH5α-pSUP at the three harvesting time points, depicted in Figure 1 .

Article Snippet: The Escherichia coli strain DH5α (F–Φ80 lac ZΔM15 Δ( lac ZYA- arg F) U169 rec A1 end A1 hsd R17 (rK−, mK+) pho A sup E44 λ− thi -1 gyr A96 rel A1) (Source: Plasmid Factory, Bielefeld, Germany) was used as a host strain for transformation of the high copy plasmid pCMV-lacZ (Source: Plasmid Factory, Bielefeld, Germany) as well as the low copy plasmid pSUP 201-3 (Simon et al., ). pCMV-lacZ is a ColE1-derived high copy plasmid for therapeutic pDNA production with a size of 7164 bp.

Techniques: Plasmid Preparation

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: ColE1-Plasmid Production in Escherichia coli : Mathematical Simulation and Experimental Validation

doi: 10.3389/fbioe.2015.00127

Figure Lengend Snippet: Measured RNAI-, RNAII-, and plasmid concentrations for DH5α-pCMV-lacZ at the three harvesting time points, depicted in Figure 2 .

Article Snippet: The Escherichia coli strain DH5α (F–Φ80 lac ZΔM15 Δ( lac ZYA- arg F) U169 rec A1 end A1 hsd R17 (rK−, mK+) pho A sup E44 λ− thi -1 gyr A96 rel A1) (Source: Plasmid Factory, Bielefeld, Germany) was used as a host strain for transformation of the high copy plasmid pCMV-lacZ (Source: Plasmid Factory, Bielefeld, Germany) as well as the low copy plasmid pSUP 201-3 (Simon et al., ). pCMV-lacZ is a ColE1-derived high copy plasmid for therapeutic pDNA production with a size of 7164 bp.

Techniques: Plasmid Preparation

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: ColE1-Plasmid Production in Escherichia coli : Mathematical Simulation and Experimental Validation

doi: 10.3389/fbioe.2015.00127

Figure Lengend Snippet: Results of the in vitro and in silico determined PCNs for DH5α-pSUP .

Article Snippet: The Escherichia coli strain DH5α (F–Φ80 lac ZΔM15 Δ( lac ZYA- arg F) U169 rec A1 end A1 hsd R17 (rK−, mK+) pho A sup E44 λ− thi -1 gyr A96 rel A1) (Source: Plasmid Factory, Bielefeld, Germany) was used as a host strain for transformation of the high copy plasmid pCMV-lacZ (Source: Plasmid Factory, Bielefeld, Germany) as well as the low copy plasmid pSUP 201-3 (Simon et al., ). pCMV-lacZ is a ColE1-derived high copy plasmid for therapeutic pDNA production with a size of 7164 bp.

Techniques: In Vitro, In Silico

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: ColE1-Plasmid Production in Escherichia coli : Mathematical Simulation and Experimental Validation

doi: 10.3389/fbioe.2015.00127

Figure Lengend Snippet: Results of the in vitro and in silico determined PCNs for DH5α-pCMV-lacZ .

Article Snippet: The Escherichia coli strain DH5α (F–Φ80 lac ZΔM15 Δ( lac ZYA- arg F) U169 rec A1 end A1 hsd R17 (rK−, mK+) pho A sup E44 λ− thi -1 gyr A96 rel A1) (Source: Plasmid Factory, Bielefeld, Germany) was used as a host strain for transformation of the high copy plasmid pCMV-lacZ (Source: Plasmid Factory, Bielefeld, Germany) as well as the low copy plasmid pSUP 201-3 (Simon et al., ). pCMV-lacZ is a ColE1-derived high copy plasmid for therapeutic pDNA production with a size of 7164 bp.

Techniques: In Vitro, In Silico

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: ColE1-Plasmid Production in Escherichia coli : Mathematical Simulation and Experimental Validation

doi: 10.3389/fbioe.2015.00127

Figure Lengend Snippet: Simulation of the plasmid replication control for E . coli DH5α-placZ, beginning at time point T 3 , for the following three different growth conditions: blue, growth with normal nutrient conditions; green, growth with amino acid starvation conditions, red, growth with modified tRNA gene without amino acid starvation conditions .

Article Snippet: The Escherichia coli strain DH5α (F–Φ80 lac ZΔM15 Δ( lac ZYA- arg F) U169 rec A1 end A1 hsd R17 (rK−, mK+) pho A sup E44 λ− thi -1 gyr A96 rel A1) (Source: Plasmid Factory, Bielefeld, Germany) was used as a host strain for transformation of the high copy plasmid pCMV-lacZ (Source: Plasmid Factory, Bielefeld, Germany) as well as the low copy plasmid pSUP 201-3 (Simon et al., ). pCMV-lacZ is a ColE1-derived high copy plasmid for therapeutic pDNA production with a size of 7164 bp.

Techniques: Plasmid Preparation, Modification