other:Article Title: Comparison of gas and liquid chromatography (GC, LC) coupled with isotope ratio mass spectrometry (IRMS): A case study on rice amino acids.
Article Snippet: The isotopic values of 9 AAs, alanine (Ala), aspartate (Asx), glutamate (Glx), glycine (Gly), isoleucine (Ile), leucine (Leu), phenylalanine (Phe), proline (Pro), and valine (Val), were determined by a Trace GC Ultra (GC IsoLink + ConFlo IV, Thermo Scientific, Bremen, Germany) interfaced with an IRMS (DELTA V, Thermo Scientific, Bremen, Germany) through an open split interface and with a single-quadrupole GC–MS (ISQ Thermo Scientific).
Recombinant:Article Title: Development of New Antimicrobial Peptides by Directional Selection
Article Snippet: .. The strain used for recombinant plasmid construction was E. coli TOP10 (F − mcrA Δ(mrr-hsdRMS-mcrBC) φ80lacZΔM15 ΔlacX74 nupG recA1 araD139 Δ(ara-leu)7697 galE15 galK16 rpsL(Str^R) endA1 λ − ) (Invitrogen, Carlsbad, CA, USA). .. The strain used for antimicrobial activity assays was E. coli BL21-Gold (DE3) (F − ompT hsdS(r_B − m_B − ) dcm + Tet^R gal λ(DE3) endA) (Novagen, Madison, WI, USA).
Plasmid Preparation:Article Title: Development of New Antimicrobial Peptides by Directional Selection
Article Snippet: .. The strain used for recombinant plasmid construction was E. coli TOP10 (F − mcrA Δ(mrr-hsdRMS-mcrBC) φ80lacZΔM15 ΔlacX74 nupG recA1 araD139 Δ(ara-leu)7697 galE15 galK16 rpsL(Str^R) endA1 λ − ) (Invitrogen, Carlsbad, CA, USA). .. The strain used for antimicrobial activity assays was E. coli BL21-Gold (DE3) (F − ompT hsdS(r_B − m_B − ) dcm + Tet^R gal λ(DE3) endA) (Novagen, Madison, WI, USA).
Article Title: Displaced by deceivers - Prevention of biosensor cross-talk is pivotal for successful biosensor-based high-throughput screening campaigns
Article Snippet: .. Strain or plasmid Relevant characteristics Reference or source E. coli DH5α F– φ80dlacZΔM15 Δ(lacZYA-argF)U169 rec A1 end A1 hsdR17 (rK–mK+) phoA supE44 thi-1 gyrA96 relA1 λ– Invitrogen E. coli TOP10 F– mcrA Δ(mrr-hsdRMS-mcrBC) φ80lacZΔM15 ΔlacX74 recA1 araD139 Δ(ara-leu) 7697 galU galK rpsL (StrR) endA1 nupG λ– Invitrogen E. coli DH10B F– mcrA Δ(mrr-hsdRMS-mcrBC) φ80lacZΔM15 ΔlacX74 recA1 endA1 araD139 Δ(ara, leu)7697 galU galK λ– rpsL nupG Invitrogen E. coli BL21(DE3) F– ompT hsdSB (rB–, mB–) gal dcm rne131 (DE3) Invitrogen E. coli DH10B ΔhcaREFCBD F– mcrA Δ(mrr-hsdRMS-mcrBC) φ80lacZΔM15 ΔlacX74 recA1 endA1 araD139 Δ(ara, leu)7697 galU galK λ– rpsL nupG ΔhcaREFCBD this work pSIJ8 Plasmid enabling L-arabinose inducible expression of λ red recombineering genes bet, gam and exo. .. L-rhamnose inducible expression of flp recombinase gene. (1) pBBR1N-reporter-EYFP Biosensor chassis plasmid with the eyfp reporter gene, KanR Markus Schallmey, unpublished pSenCA Transcriptional biosensor construct inducing eyfp expression in response to the presence of trans-cinnamic acid or phenylpropionic acid, KanR This work pBAD/myc-His A Cloning vector for the synthesis of His6-tagged (c-terminal) fusion protein, AmpR Invitrogen pCBJ296 pCDFDuet-1 MCS2::xalTc, SpR (2) pBAD-xalTc Cloning vector for XalTc protein synthesis, AmpR This work pBAD-N6XHIS Cloning vector for the synthesis of His6-tagged (n-terminal) protein, AmpR This work pBAD-N6XHIS-xalTc Cloning vector for the synthesis of His6-tagged (n-terminal) XalTc fusion protein, AmpR This work Supplementary Table S2.
Article Title: Enhancing Protein Stability with Genetically Encoded Noncanonical Amino Acids
Article Snippet: Single peaks were extracted and deconvoluted using Agilent Masshunter software. .. Strains and plasmids used Strains and plasmids Description Source E. coli DH10B F- mcrA Δ(mrr-hsdRMS-mcrBC) Φ80lacZΔM15 lacX74 recA1 endA1 araD139 Δ(ara leu) 7697 galU galK rpsL nupG λ– Thermo Fisher E. coli JW3973-1 F-, Δ(araD-araB)567, ΔlacZ4787(::rrnB-3), λ-, rph-1, Δ(rhaDrhaB)568, ΔmetA780::kan, hsdR514 Yale CGSC pUltra-polyRS Plasmid containing polyspecific tyrosine analog synthetase/tRNA pair CloDF13 ori, Smr, tac promoter, IPTG inducible synthetase expression, constitutive tRNA expression Schultz laboratory 1 pEVOL-pBzF Plasmid containing synthetase/tRNA pair for p-benzoyl phenylalanine p15A ori, catr, arabinose inducible and constitutive synthetase expression, constitutive tRNA expression Schultz laboratory 2 pEVOL-pBoF Plasmid containing synthetase/tRNA pair for p-borono phenylalanine p15A ori, catr, arabinose inducible and constitutive synthetase expression, constitutive tRNA expression Schultz laboratory 3 pUltra-VSF Plasmid containing synthetase/tRNA pair for p-vinylsulfamido phenylalanine CloDF13 ori, Smr, tac promoter, IPTG inducible synthetase expression, constitutive tRNA expression Schultz laboratory 4 pBR322 metA metA plasmid used for library construction and screening pBR322 ori, Ampr, constitutively expressing metA This study pBAD metA Plasmid used for his tagged N-terminal 6xHis metA overexpression. .. Strains and plasmids used Strains and plasmids Description Source E. coli DH10B F- mcrA Δ(mrr-hsdRMS-mcrBC) Φ80lacZΔM15 lacX74 recA1 endA1 araD139 Δ(ara leu) 7697 galU galK rpsL nupG λ– Thermo Fisher E. coli JW3973-1 F-, Δ(araD-araB)567, ΔlacZ4787(::rrnB-3), λ-, rph-1, Δ(rhaDrhaB)568, ΔmetA780::kan, hsdR514 Yale CGSC pUltra-polyRS Plasmid containing polyspecific tyrosine analog synthetase/tRNA pair CloDF13 ori, Smr, tac promoter, IPTG inducible synthetase expression, constitutive tRNA expression Schultz laboratory 1 pEVOL-pBzF Plasmid containing synthetase/tRNA pair for p-benzoyl phenylalanine p15A ori, catr, arabinose inducible and constitutive synthetase expression, constitutive tRNA expression Schultz laboratory 2 pEVOL-pBoF Plasmid containing synthetase/tRNA pair for p-borono phenylalanine p15A ori, catr, arabinose inducible and constitutive synthetase expression, constitutive tRNA expression Schultz laboratory 3 pUltra-VSF Plasmid containing synthetase/tRNA pair for p-vinylsulfamido phenylalanine CloDF13 ori, Smr, tac promoter, IPTG inducible synthetase expression, constitutive tRNA expression Schultz laboratory 4 pBR322 metA metA plasmid used for library construction and screening pBR322 ori, Ampr, constitutively expressing metA This study pBAD metA Plasmid used for his tagged N-terminal 6xHis metA overexpression.
Expressing:Article Title: Displaced by deceivers - Prevention of biosensor cross-talk is pivotal for successful biosensor-based high-throughput screening campaigns
Article Snippet: .. Strain or plasmid Relevant characteristics Reference or source E. coli DH5α F– φ80dlacZΔM15 Δ(lacZYA-argF)U169 rec A1 end A1 hsdR17 (rK–mK+) phoA supE44 thi-1 gyrA96 relA1 λ– Invitrogen E. coli TOP10 F– mcrA Δ(mrr-hsdRMS-mcrBC) φ80lacZΔM15 ΔlacX74 recA1 araD139 Δ(ara-leu) 7697 galU galK rpsL (StrR) endA1 nupG λ– Invitrogen E. coli DH10B F– mcrA Δ(mrr-hsdRMS-mcrBC) φ80lacZΔM15 ΔlacX74 recA1 endA1 araD139 Δ(ara, leu)7697 galU galK λ– rpsL nupG Invitrogen E. coli BL21(DE3) F– ompT hsdSB (rB–, mB–) gal dcm rne131 (DE3) Invitrogen E. coli DH10B ΔhcaREFCBD F– mcrA Δ(mrr-hsdRMS-mcrBC) φ80lacZΔM15 ΔlacX74 recA1 endA1 araD139 Δ(ara, leu)7697 galU galK λ– rpsL nupG ΔhcaREFCBD this work pSIJ8 Plasmid enabling L-arabinose inducible expression of λ red recombineering genes bet, gam and exo. .. L-rhamnose inducible expression of flp recombinase gene. (1) pBBR1N-reporter-EYFP Biosensor chassis plasmid with the eyfp reporter gene, KanR Markus Schallmey, unpublished pSenCA Transcriptional biosensor construct inducing eyfp expression in response to the presence of trans-cinnamic acid or phenylpropionic acid, KanR This work pBAD/myc-His A Cloning vector for the synthesis of His6-tagged (c-terminal) fusion protein, AmpR Invitrogen pCBJ296 pCDFDuet-1 MCS2::xalTc, SpR (2) pBAD-xalTc Cloning vector for XalTc protein synthesis, AmpR This work pBAD-N6XHIS Cloning vector for the synthesis of His6-tagged (n-terminal) protein, AmpR This work pBAD-N6XHIS-xalTc Cloning vector for the synthesis of His6-tagged (n-terminal) XalTc fusion protein, AmpR This work Supplementary Table S2.
Article Title: Enhancing Protein Stability with Genetically Encoded Noncanonical Amino Acids
Article Snippet: Single peaks were extracted and deconvoluted using Agilent Masshunter software. .. Strains and plasmids used Strains and plasmids Description Source E. coli DH10B F- mcrA Δ(mrr-hsdRMS-mcrBC) Φ80lacZΔM15 lacX74 recA1 endA1 araD139 Δ(ara leu) 7697 galU galK rpsL nupG λ– Thermo Fisher E. coli JW3973-1 F-, Δ(araD-araB)567, ΔlacZ4787(::rrnB-3), λ-, rph-1, Δ(rhaDrhaB)568, ΔmetA780::kan, hsdR514 Yale CGSC pUltra-polyRS Plasmid containing polyspecific tyrosine analog synthetase/tRNA pair CloDF13 ori, Smr, tac promoter, IPTG inducible synthetase expression, constitutive tRNA expression Schultz laboratory 1 pEVOL-pBzF Plasmid containing synthetase/tRNA pair for p-benzoyl phenylalanine p15A ori, catr, arabinose inducible and constitutive synthetase expression, constitutive tRNA expression Schultz laboratory 2 pEVOL-pBoF Plasmid containing synthetase/tRNA pair for p-borono phenylalanine p15A ori, catr, arabinose inducible and constitutive synthetase expression, constitutive tRNA expression Schultz laboratory 3 pUltra-VSF Plasmid containing synthetase/tRNA pair for p-vinylsulfamido phenylalanine CloDF13 ori, Smr, tac promoter, IPTG inducible synthetase expression, constitutive tRNA expression Schultz laboratory 4 pBR322 metA metA plasmid used for library construction and screening pBR322 ori, Ampr, constitutively expressing metA This study pBAD metA Plasmid used for his tagged N-terminal 6xHis metA overexpression. .. Strains and plasmids used Strains and plasmids Description Source E. coli DH10B F- mcrA Δ(mrr-hsdRMS-mcrBC) Φ80lacZΔM15 lacX74 recA1 endA1 araD139 Δ(ara leu) 7697 galU galK rpsL nupG λ– Thermo Fisher E. coli JW3973-1 F-, Δ(araD-araB)567, ΔlacZ4787(::rrnB-3), λ-, rph-1, Δ(rhaDrhaB)568, ΔmetA780::kan, hsdR514 Yale CGSC pUltra-polyRS Plasmid containing polyspecific tyrosine analog synthetase/tRNA pair CloDF13 ori, Smr, tac promoter, IPTG inducible synthetase expression, constitutive tRNA expression Schultz laboratory 1 pEVOL-pBzF Plasmid containing synthetase/tRNA pair for p-benzoyl phenylalanine p15A ori, catr, arabinose inducible and constitutive synthetase expression, constitutive tRNA expression Schultz laboratory 2 pEVOL-pBoF Plasmid containing synthetase/tRNA pair for p-borono phenylalanine p15A ori, catr, arabinose inducible and constitutive synthetase expression, constitutive tRNA expression Schultz laboratory 3 pUltra-VSF Plasmid containing synthetase/tRNA pair for p-vinylsulfamido phenylalanine CloDF13 ori, Smr, tac promoter, IPTG inducible synthetase expression, constitutive tRNA expression Schultz laboratory 4 pBR322 metA metA plasmid used for library construction and screening pBR322 ori, Ampr, constitutively expressing metA This study pBAD metA Plasmid used for his tagged N-terminal 6xHis metA overexpression.
Incubation:Article Title: GATA2 decommissioning of enhancers: A mechanism to constrain inflammatory signaling.
Article Snippet: Total RNA was purified from 1-5 × 105 cells with TRIzol (Invitrogen) and 2 μg RNA was treated with DNase I (Thermo Fisher Scientific) for 15 min at room temperature. .. After heat inactivation of DNase I with EDTA for 10 min at 65◦C, RNA was incubated with 250 ng of a 4:1 mixture of Oligo dT primers and Random Hexamer at 68◦C for 10 min. RNA/primers were incubated with Moloney Murine Leu- kemia Virus Reverse Transcriptase (M-MLV RT) (Thermo Fisher Scientific), 5× First Strand Buffer (Thermo Fisher Scientific), 10 mM Dithiothreitol (Thermo Fisher Scientific), RNAsin (Promega), and 0.5 mM Deoxynucleoside Triphosphates (New England Biolabs) at 42◦C for 1 h and then heat inactivated at 95◦C for 5 min. Quantitative gene expression analysis was performed by real-time RT-qPCR using Power SYBR Green Master Mix (Applied Biosystems) and analyzed on a ViiA 7 Real-Time PCR System (Applied Biosystems). ..
Article Title: PGC-1α and PPARs cooperatively mediate photoreceptor neuroprotection in rd1 mouse inherited retinal degeneration
Article Snippet: Finally, sections were washed three times with PBS (5 min each), mounted in Vectashield with DAPI (Vector Laboratories Inc., Burlingame, CA, USA), and imaged by microscopy. .. Unfixed retinal tissue sections were rehydrated in calpain reaction buffer (CRB) (5.96 g HEPES, 4.85 g KCl, 0.47 g MgCl2, and 0.22 g CaCl2 in 100 mL ddH2O; pH 7.2) with 2 mM dithiothreitol (DTT) for 15 min. Then, sections were incubated with 25 μM tBOC-Leu-Met-CMAC (A6520; Thermo Fisher Scientific, OR, USA) for 3 h at 37 °C. .. This was followed by two washes with PBS and 12 min incubation with ToPro3 (1:1000 in PBS; T3605, Invitrogen, Carlsbad, USA).
Random Hexamer:Article Title: GATA2 decommissioning of enhancers: A mechanism to constrain inflammatory signaling.
Article Snippet: Total RNA was purified from 1-5 × 105 cells with TRIzol (Invitrogen) and 2 μg RNA was treated with DNase I (Thermo Fisher Scientific) for 15 min at room temperature. .. After heat inactivation of DNase I with EDTA for 10 min at 65◦C, RNA was incubated with 250 ng of a 4:1 mixture of Oligo dT primers and Random Hexamer at 68◦C for 10 min. RNA/primers were incubated with Moloney Murine Leu- kemia Virus Reverse Transcriptase (M-MLV RT) (Thermo Fisher Scientific), 5× First Strand Buffer (Thermo Fisher Scientific), 10 mM Dithiothreitol (Thermo Fisher Scientific), RNAsin (Promega), and 0.5 mM Deoxynucleoside Triphosphates (New England Biolabs) at 42◦C for 1 h and then heat inactivated at 95◦C for 5 min. Quantitative gene expression analysis was performed by real-time RT-qPCR using Power SYBR Green Master Mix (Applied Biosystems) and analyzed on a ViiA 7 Real-Time PCR System (Applied Biosystems). ..
Virus:Article Title: GATA2 decommissioning of enhancers: A mechanism to constrain inflammatory signaling.
Article Snippet: Total RNA was purified from 1-5 × 105 cells with TRIzol (Invitrogen) and 2 μg RNA was treated with DNase I (Thermo Fisher Scientific) for 15 min at room temperature. .. After heat inactivation of DNase I with EDTA for 10 min at 65◦C, RNA was incubated with 250 ng of a 4:1 mixture of Oligo dT primers and Random Hexamer at 68◦C for 10 min. RNA/primers were incubated with Moloney Murine Leu- kemia Virus Reverse Transcriptase (M-MLV RT) (Thermo Fisher Scientific), 5× First Strand Buffer (Thermo Fisher Scientific), 10 mM Dithiothreitol (Thermo Fisher Scientific), RNAsin (Promega), and 0.5 mM Deoxynucleoside Triphosphates (New England Biolabs) at 42◦C for 1 h and then heat inactivated at 95◦C for 5 min. Quantitative gene expression analysis was performed by real-time RT-qPCR using Power SYBR Green Master Mix (Applied Biosystems) and analyzed on a ViiA 7 Real-Time PCR System (Applied Biosystems). ..
Reverse Transcription:Article Title: GATA2 decommissioning of enhancers: A mechanism to constrain inflammatory signaling.
Article Snippet: Total RNA was purified from 1-5 × 105 cells with TRIzol (Invitrogen) and 2 μg RNA was treated with DNase I (Thermo Fisher Scientific) for 15 min at room temperature. .. After heat inactivation of DNase I with EDTA for 10 min at 65◦C, RNA was incubated with 250 ng of a 4:1 mixture of Oligo dT primers and Random Hexamer at 68◦C for 10 min. RNA/primers were incubated with Moloney Murine Leu- kemia Virus Reverse Transcriptase (M-MLV RT) (Thermo Fisher Scientific), 5× First Strand Buffer (Thermo Fisher Scientific), 10 mM Dithiothreitol (Thermo Fisher Scientific), RNAsin (Promega), and 0.5 mM Deoxynucleoside Triphosphates (New England Biolabs) at 42◦C for 1 h and then heat inactivated at 95◦C for 5 min. Quantitative gene expression analysis was performed by real-time RT-qPCR using Power SYBR Green Master Mix (Applied Biosystems) and analyzed on a ViiA 7 Real-Time PCR System (Applied Biosystems). ..
Gene Expression:Article Title: GATA2 decommissioning of enhancers: A mechanism to constrain inflammatory signaling.
Article Snippet: Total RNA was purified from 1-5 × 105 cells with TRIzol (Invitrogen) and 2 μg RNA was treated with DNase I (Thermo Fisher Scientific) for 15 min at room temperature. .. After heat inactivation of DNase I with EDTA for 10 min at 65◦C, RNA was incubated with 250 ng of a 4:1 mixture of Oligo dT primers and Random Hexamer at 68◦C for 10 min. RNA/primers were incubated with Moloney Murine Leu- kemia Virus Reverse Transcriptase (M-MLV RT) (Thermo Fisher Scientific), 5× First Strand Buffer (Thermo Fisher Scientific), 10 mM Dithiothreitol (Thermo Fisher Scientific), RNAsin (Promega), and 0.5 mM Deoxynucleoside Triphosphates (New England Biolabs) at 42◦C for 1 h and then heat inactivated at 95◦C for 5 min. Quantitative gene expression analysis was performed by real-time RT-qPCR using Power SYBR Green Master Mix (Applied Biosystems) and analyzed on a ViiA 7 Real-Time PCR System (Applied Biosystems). ..
Quantitative RT-PCR:Article Title: GATA2 decommissioning of enhancers: A mechanism to constrain inflammatory signaling.
Article Snippet: Total RNA was purified from 1-5 × 105 cells with TRIzol (Invitrogen) and 2 μg RNA was treated with DNase I (Thermo Fisher Scientific) for 15 min at room temperature. .. After heat inactivation of DNase I with EDTA for 10 min at 65◦C, RNA was incubated with 250 ng of a 4:1 mixture of Oligo dT primers and Random Hexamer at 68◦C for 10 min. RNA/primers were incubated with Moloney Murine Leu- kemia Virus Reverse Transcriptase (M-MLV RT) (Thermo Fisher Scientific), 5× First Strand Buffer (Thermo Fisher Scientific), 10 mM Dithiothreitol (Thermo Fisher Scientific), RNAsin (Promega), and 0.5 mM Deoxynucleoside Triphosphates (New England Biolabs) at 42◦C for 1 h and then heat inactivated at 95◦C for 5 min. Quantitative gene expression analysis was performed by real-time RT-qPCR using Power SYBR Green Master Mix (Applied Biosystems) and analyzed on a ViiA 7 Real-Time PCR System (Applied Biosystems). ..
SYBR Green Assay:Article Title: GATA2 decommissioning of enhancers: A mechanism to constrain inflammatory signaling.
Article Snippet: Total RNA was purified from 1-5 × 105 cells with TRIzol (Invitrogen) and 2 μg RNA was treated with DNase I (Thermo Fisher Scientific) for 15 min at room temperature. .. After heat inactivation of DNase I with EDTA for 10 min at 65◦C, RNA was incubated with 250 ng of a 4:1 mixture of Oligo dT primers and Random Hexamer at 68◦C for 10 min. RNA/primers were incubated with Moloney Murine Leu- kemia Virus Reverse Transcriptase (M-MLV RT) (Thermo Fisher Scientific), 5× First Strand Buffer (Thermo Fisher Scientific), 10 mM Dithiothreitol (Thermo Fisher Scientific), RNAsin (Promega), and 0.5 mM Deoxynucleoside Triphosphates (New England Biolabs) at 42◦C for 1 h and then heat inactivated at 95◦C for 5 min. Quantitative gene expression analysis was performed by real-time RT-qPCR using Power SYBR Green Master Mix (Applied Biosystems) and analyzed on a ViiA 7 Real-Time PCR System (Applied Biosystems). ..
Real-time Polymerase Chain Reaction:Article Title: GATA2 decommissioning of enhancers: A mechanism to constrain inflammatory signaling.
Article Snippet: Total RNA was purified from 1-5 × 105 cells with TRIzol (Invitrogen) and 2 μg RNA was treated with DNase I (Thermo Fisher Scientific) for 15 min at room temperature. .. After heat inactivation of DNase I with EDTA for 10 min at 65◦C, RNA was incubated with 250 ng of a 4:1 mixture of Oligo dT primers and Random Hexamer at 68◦C for 10 min. RNA/primers were incubated with Moloney Murine Leu- kemia Virus Reverse Transcriptase (M-MLV RT) (Thermo Fisher Scientific), 5× First Strand Buffer (Thermo Fisher Scientific), 10 mM Dithiothreitol (Thermo Fisher Scientific), RNAsin (Promega), and 0.5 mM Deoxynucleoside Triphosphates (New England Biolabs) at 42◦C for 1 h and then heat inactivated at 95◦C for 5 min. Quantitative gene expression analysis was performed by real-time RT-qPCR using Power SYBR Green Master Mix (Applied Biosystems) and analyzed on a ViiA 7 Real-Time PCR System (Applied Biosystems). ..
Construct:Article Title: Substituted 2,4-dioxotetrahydropyrimidines as intermediates in the synthesis of bruton's tyrosine kinase inhibitors
Article Snippet: For C-terminal tagging the STOP codon was mutated to a leucine performing a mutagenesis reaction with the following primers (pENTR221-BTK Quikchange STOP-Leu fw, gtcatggatgaagaatccTTGaacccagctttcttgtac; pENTR221-BTK Quikchange STOP-Leu REVC, gtacaagaaagctgggttCAAggattcttcatccatgac) using the QuikChange Lightning mutagenesis kit (Agilent Technologies #210518) according to the manufacturer's manual, resulting in pENTR221-BTK (STOP-Leu). .. To get the final pLenti6-BTK-GFP-Ires-RFP sensor construct, a Gateway LR reaction was performed between pLenti6-DEST-GFP-lres-RFP and pENTR221-BTK (STOP-Leu) using the LR Clonase kit (Invitrogen 11791-019) according to the manufacturer's manual. ..
Over Expression:Article Title: Enhancing Protein Stability with Genetically Encoded Noncanonical Amino Acids
Article Snippet: Single peaks were extracted and deconvoluted using Agilent Masshunter software. .. Strains and plasmids used Strains and plasmids Description Source E. coli DH10B F- mcrA Δ(mrr-hsdRMS-mcrBC) Φ80lacZΔM15 lacX74 recA1 endA1 araD139 Δ(ara leu) 7697 galU galK rpsL nupG λ– Thermo Fisher E. coli JW3973-1 F-, Δ(araD-araB)567, ΔlacZ4787(::rrnB-3), λ-, rph-1, Δ(rhaDrhaB)568, ΔmetA780::kan, hsdR514 Yale CGSC pUltra-polyRS Plasmid containing polyspecific tyrosine analog synthetase/tRNA pair CloDF13 ori, Smr, tac promoter, IPTG inducible synthetase expression, constitutive tRNA expression Schultz laboratory 1 pEVOL-pBzF Plasmid containing synthetase/tRNA pair for p-benzoyl phenylalanine p15A ori, catr, arabinose inducible and constitutive synthetase expression, constitutive tRNA expression Schultz laboratory 2 pEVOL-pBoF Plasmid containing synthetase/tRNA pair for p-borono phenylalanine p15A ori, catr, arabinose inducible and constitutive synthetase expression, constitutive tRNA expression Schultz laboratory 3 pUltra-VSF Plasmid containing synthetase/tRNA pair for p-vinylsulfamido phenylalanine CloDF13 ori, Smr, tac promoter, IPTG inducible synthetase expression, constitutive tRNA expression Schultz laboratory 4 pBR322 metA metA plasmid used for library construction and screening pBR322 ori, Ampr, constitutively expressing metA This study pBAD metA Plasmid used for his tagged N-terminal 6xHis metA overexpression. .. Strains and plasmids used Strains and plasmids Description Source E. coli DH10B F- mcrA Δ(mrr-hsdRMS-mcrBC) Φ80lacZΔM15 lacX74 recA1 endA1 araD139 Δ(ara leu) 7697 galU galK rpsL nupG λ– Thermo Fisher E. coli JW3973-1 F-, Δ(araD-araB)567, ΔlacZ4787(::rrnB-3), λ-, rph-1, Δ(rhaDrhaB)568, ΔmetA780::kan, hsdR514 Yale CGSC pUltra-polyRS Plasmid containing polyspecific tyrosine analog synthetase/tRNA pair CloDF13 ori, Smr, tac promoter, IPTG inducible synthetase expression, constitutive tRNA expression Schultz laboratory 1 pEVOL-pBzF Plasmid containing synthetase/tRNA pair for p-benzoyl phenylalanine p15A ori, catr, arabinose inducible and constitutive synthetase expression, constitutive tRNA expression Schultz laboratory 2 pEVOL-pBoF Plasmid containing synthetase/tRNA pair for p-borono phenylalanine p15A ori, catr, arabinose inducible and constitutive synthetase expression, constitutive tRNA expression Schultz laboratory 3 pUltra-VSF Plasmid containing synthetase/tRNA pair for p-vinylsulfamido phenylalanine CloDF13 ori, Smr, tac promoter, IPTG inducible synthetase expression, constitutive tRNA expression Schultz laboratory 4 pBR322 metA metA plasmid used for library construction and screening pBR322 ori, Ampr, constitutively expressing metA This study pBAD metA Plasmid used for his tagged N-terminal 6xHis metA overexpression.
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