e coli top10 e coli cloning strain  (New England Biolabs)

Journal: Genes

Article Title: Modulating the Precursor and Terpene Synthase Supply for the Whole-Cell Biocatalytic Production of the Sesquiterpene (+)-Zizaene in a Pathway Engineered E. coli

doi: 10.3390/genes10060478

Figure Lengend Snippet: Strains and plasmids used in this investigation.

Article Snippet: For that, both the insert and backbone were amplified with complementary overhangs by PCR methods, seamlessly assembled, and E. coli TOP10 competent cloning strains (IBA, Germany) were transformed for respective screening.

Techniques: Plasmid Preparation

Journal: Genes

Article Title: Modulating the Precursor and Terpene Synthase Supply for the Whole-Cell Biocatalytic Production of the Sesquiterpene (+)-Zizaene in a Pathway Engineered E. coli

doi: 10.3390/genes10060478

Figure Lengend Snippet: Biosynthetic pathway engineered for the production of (+)-zizaene in E. coli by the expression of the native methylerythritol phosphate (MEP) pathway, the heterologous MEV pathway and the (+)-zizaene pathway. Heterologous expressed enzymes highlighted in blue. The MEP pathway comprise the following enzymes: 1-Deoxy-D-xylulose 5-phosphate synthase ( Dxs ), 1-deoxy-D-xylulose 5-phosphate reductoisomerase ( Dxr ), MEP cytidyl transferase ( MCT ), cytidyl MEP kinase ( CMK ), MEP-2,4-cyclodiphosphate synthase ( MDS ), (E)-4-hydroxy-3-methylbut-2-enyl diphosphate synthase ( HDS ), and (E)-4-hydroxy-3-methylbut-2-enyl diphosphate reductase ( HDR ). The MEV heterologous pathway consisted of the following enzymes: acetyl-CoA acetyltransferase ( AtoB ), HMG-CoA synthase (HMGS), truncated HMG-CoA reductase ( tHMGR ), mevalonate kinase (MK), phosphomevalonate kinase (PMK), mevalonate diphosphate decarboxylase (PMD), and isoprenyl diphosphate isomerase ( Idi ). The (+)-zizaene pathway comprises the farnesyl diphosphate synthase ( IspA ) and the zizaene synthase ( ZS ).

Article Snippet: For that, both the insert and backbone were amplified with complementary overhangs by PCR methods, seamlessly assembled, and E. coli TOP10 competent cloning strains (IBA, Germany) were transformed for respective screening.

Techniques: Expressing

Journal: Genes

Article Title: Modulating the Precursor and Terpene Synthase Supply for the Whole-Cell Biocatalytic Production of the Sesquiterpene (+)-Zizaene in a Pathway Engineered E. coli

doi: 10.3390/genes10060478

Figure Lengend Snippet: Comparison of the performance of the engineered E. coli BL21(DE3) strains induced at different IPTG levels. ( A ) (+)-zizaene titers. ( B ) (+)-zizaene yields. ( C ) biomass from cell dry weight ( D ) cell density. Data are the mean of four independent replicates from production cultures after 24 h of induction with DNB medium, and the error bars represent the standard deviation. * BZS+Mev E. coli Top10 strain induced with 1 mM IPTG and different levels of arabinose (Ara).

Article Snippet: For that, both the insert and backbone were amplified with complementary overhangs by PCR methods, seamlessly assembled, and E. coli TOP10 competent cloning strains (IBA, Germany) were transformed for respective screening.

Techniques: Standard Deviation

Journal: Genes

Article Title: Modulating the Precursor and Terpene Synthase Supply for the Whole-Cell Biocatalytic Production of the Sesquiterpene (+)-Zizaene in a Pathway Engineered E. coli

doi: 10.3390/genes10060478

Figure Lengend Snippet: Evaluation of the overexpression of the soluble (SF) and insoluble (IF) ZS protein fractions from the engineered E. coli BL21(DE3) strains, induced at different IPTG levels by 10% SDS-PAGE. (M) molecular marker. * BZS+Mev induced with 1 mM IPTG and different levels of arabinose (Ara).

Article Snippet: For that, both the insert and backbone were amplified with complementary overhangs by PCR methods, seamlessly assembled, and E. coli TOP10 competent cloning strains (IBA, Germany) were transformed for respective screening.

Techniques: Over Expression, SDS Page, Marker

Journal: Genes

Article Title: Modulating the Precursor and Terpene Synthase Supply for the Whole-Cell Biocatalytic Production of the Sesquiterpene (+)-Zizaene in a Pathway Engineered E. coli

doi: 10.3390/genes10060478

Figure Lengend Snippet: Optimization of the fermentation conditions and testing of E. coli strains for the improvement of the terpene performance. Evaluation of production media: ( A ) (+)-zizaene titers and yields, ( B ) biomass and cell density. Optimization of growth temperature and media pH: ( C ) (+)-zizaene titers, ( D ) (+)-zizaene yields. Evaluation of E. coli strains: ( E ) (+)-zizaene titers and yields, ( F ) biomass and cell density. Data are the mean of four independent replicates from production cultures after 24 h of induction, and the error bars represent the standard deviation.

Article Snippet: For that, both the insert and backbone were amplified with complementary overhangs by PCR methods, seamlessly assembled, and E. coli TOP10 competent cloning strains (IBA, Germany) were transformed for respective screening.

Techniques: Standard Deviation

Journal: Genes

Article Title: Modulating the Precursor and Terpene Synthase Supply for the Whole-Cell Biocatalytic Production of the Sesquiterpene (+)-Zizaene in a Pathway Engineered E. coli

doi: 10.3390/genes10060478

Figure Lengend Snippet: Analysis of the over-expression of soluble (SF) and insoluble (IF) ZS protein fractions from the optimization of fermentation conditions and testing of E. coli strains by 10% SDS-PAGE. ( A ) Culture media test. ( B ) Temperature and pH optimization. ( C ) E. coli strains test. (M) molecular marker.

Article Snippet: For that, both the insert and backbone were amplified with complementary overhangs by PCR methods, seamlessly assembled, and E. coli TOP10 competent cloning strains (IBA, Germany) were transformed for respective screening.

Techniques: Over Expression, SDS Page, Marker