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    GeneArt Strings DNA Libraries randomized bp
    Description:
    GeneArt Strings DNA Fragments are custom made uncloned double stranded linear DNA fragments from 150 base pairs bp to 3 000 bp in length assembled from synthetic oligonucleotides using the same process developed for GeneArt high quality gene synthesis
    Catalog Number:
    833007de
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    DNA Vectors Clones Purified Nucleic Acids Libraries
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    Structured Review

    Thermo Fisher strings dna fragments
    Characterization of a light-gated potassium channel in hippocampal neurons. (A) Maximum intensity projection of confocal images of hippocampal neurons 3 days after electroporation with <t>DNA</t> encoding <t>SthK-bP</t> and mKate2 (left: excitation 559 nm; right: excitation 488 nm). Lower images are close up of the regions indicated by dark boxes. (B) Single plane of the YFP signal of the region indicated in (A) (dark yellow box). (C) Left: Sample photocurrents evoked by a 50 ms light pulse (470 nm) of different intensity in a hippocampal neuron expressing mKate2 and SthK-bP. The holding potential was −70 mV. Right: Light intensity-response relationship fitted with a quadratic equation. Photocurrents are normalized to the maximum current recorded in each neuron. n = 5. (D) Left: Sample traces of photocurrents recorded from a SthK-bP expressing neuron when stimulated with 0.1 mW/mm 2 , 470 nm light (50 ms) while holding the cell at membrane potentials from −70 mV down to −115 mV. Baselines are aligned. Right: Current-voltage plot. A non-linear fit was applied to determine the K + equilibrium potential (−100.5 mV). (E) Photocurrent amplitude recorded from neurons expressing mKate2 and SthK-bP when stimulated with 1 mW mm −2 470 nm light. Shown are individual data points, median and 25–75% interquartile range, n = 5; median peak current SthK-bP 1.37 nA. (F) Kinetics of SthK-bP; data obtained from traces when the stimulation intensity was 1 mW mm −2 (470 nm, 50 ms); n = 5. (G) Whole-cell responses to current injections from −400 to 700 pA in SthK-bP expressing hippocampal neuron. (H) Action potentials generated by repeated somatic current injection (1,000 pA, 600 ms, ISI 5 s) were blocked for 3 min by 3 470 nm light flashes at 40 second intervals (50 ms at 1 mW mm −2 ); same neuron as in (G) .
    GeneArt Strings DNA Fragments are custom made uncloned double stranded linear DNA fragments from 150 base pairs bp to 3 000 bp in length assembled from synthetic oligonucleotides using the same process developed for GeneArt high quality gene synthesis
    https://www.bioz.com/result/strings dna fragments/product/Thermo Fisher
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    strings dna fragments - by Bioz Stars, 2021-03
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    Images

    1) Product Images from "Synthetic Light-Activated Ion Channels for Optogenetic Activation and Inhibition"

    Article Title: Synthetic Light-Activated Ion Channels for Optogenetic Activation and Inhibition

    Journal: Frontiers in Neuroscience

    doi: 10.3389/fnins.2018.00643

    Characterization of a light-gated potassium channel in hippocampal neurons. (A) Maximum intensity projection of confocal images of hippocampal neurons 3 days after electroporation with DNA encoding SthK-bP and mKate2 (left: excitation 559 nm; right: excitation 488 nm). Lower images are close up of the regions indicated by dark boxes. (B) Single plane of the YFP signal of the region indicated in (A) (dark yellow box). (C) Left: Sample photocurrents evoked by a 50 ms light pulse (470 nm) of different intensity in a hippocampal neuron expressing mKate2 and SthK-bP. The holding potential was −70 mV. Right: Light intensity-response relationship fitted with a quadratic equation. Photocurrents are normalized to the maximum current recorded in each neuron. n = 5. (D) Left: Sample traces of photocurrents recorded from a SthK-bP expressing neuron when stimulated with 0.1 mW/mm 2 , 470 nm light (50 ms) while holding the cell at membrane potentials from −70 mV down to −115 mV. Baselines are aligned. Right: Current-voltage plot. A non-linear fit was applied to determine the K + equilibrium potential (−100.5 mV). (E) Photocurrent amplitude recorded from neurons expressing mKate2 and SthK-bP when stimulated with 1 mW mm −2 470 nm light. Shown are individual data points, median and 25–75% interquartile range, n = 5; median peak current SthK-bP 1.37 nA. (F) Kinetics of SthK-bP; data obtained from traces when the stimulation intensity was 1 mW mm −2 (470 nm, 50 ms); n = 5. (G) Whole-cell responses to current injections from −400 to 700 pA in SthK-bP expressing hippocampal neuron. (H) Action potentials generated by repeated somatic current injection (1,000 pA, 600 ms, ISI 5 s) were blocked for 3 min by 3 470 nm light flashes at 40 second intervals (50 ms at 1 mW mm −2 ); same neuron as in (G) .
    Figure Legend Snippet: Characterization of a light-gated potassium channel in hippocampal neurons. (A) Maximum intensity projection of confocal images of hippocampal neurons 3 days after electroporation with DNA encoding SthK-bP and mKate2 (left: excitation 559 nm; right: excitation 488 nm). Lower images are close up of the regions indicated by dark boxes. (B) Single plane of the YFP signal of the region indicated in (A) (dark yellow box). (C) Left: Sample photocurrents evoked by a 50 ms light pulse (470 nm) of different intensity in a hippocampal neuron expressing mKate2 and SthK-bP. The holding potential was −70 mV. Right: Light intensity-response relationship fitted with a quadratic equation. Photocurrents are normalized to the maximum current recorded in each neuron. n = 5. (D) Left: Sample traces of photocurrents recorded from a SthK-bP expressing neuron when stimulated with 0.1 mW/mm 2 , 470 nm light (50 ms) while holding the cell at membrane potentials from −70 mV down to −115 mV. Baselines are aligned. Right: Current-voltage plot. A non-linear fit was applied to determine the K + equilibrium potential (−100.5 mV). (E) Photocurrent amplitude recorded from neurons expressing mKate2 and SthK-bP when stimulated with 1 mW mm −2 470 nm light. Shown are individual data points, median and 25–75% interquartile range, n = 5; median peak current SthK-bP 1.37 nA. (F) Kinetics of SthK-bP; data obtained from traces when the stimulation intensity was 1 mW mm −2 (470 nm, 50 ms); n = 5. (G) Whole-cell responses to current injections from −400 to 700 pA in SthK-bP expressing hippocampal neuron. (H) Action potentials generated by repeated somatic current injection (1,000 pA, 600 ms, ISI 5 s) were blocked for 3 min by 3 470 nm light flashes at 40 second intervals (50 ms at 1 mW mm −2 ); same neuron as in (G) .

    Techniques Used: Electroporation, Mass Spectrometry, Expressing, Generated, Injection

    2) Product Images from "Synthetic Light-Activated Ion Channels for Optogenetic Activation and Inhibition"

    Article Title: Synthetic Light-Activated Ion Channels for Optogenetic Activation and Inhibition

    Journal: Frontiers in Neuroscience

    doi: 10.3389/fnins.2018.00643

    Characterization of a light-gated potassium channel in hippocampal neurons. (A) Maximum intensity projection of confocal images of hippocampal neurons 3 days after electroporation with DNA encoding SthK-bP and mKate2 (left: excitation 559 nm; right: excitation 488 nm). Lower images are close up of the regions indicated by dark boxes. (B) Single plane of the YFP signal of the region indicated in (A) (dark yellow box). (C) Left: Sample photocurrents evoked by a 50 ms light pulse (470 nm) of different intensity in a hippocampal neuron expressing mKate2 and SthK-bP. The holding potential was −70 mV. Right: Light intensity-response relationship fitted with a quadratic equation. Photocurrents are normalized to the maximum current recorded in each neuron. n = 5. (D) Left: Sample traces of photocurrents recorded from a SthK-bP expressing neuron when stimulated with 0.1 mW/mm 2 , 470 nm light (50 ms) while holding the cell at membrane potentials from −70 mV down to −115 mV. Baselines are aligned. Right: Current-voltage plot. A non-linear fit was applied to determine the K + equilibrium potential (−100.5 mV). (E) Photocurrent amplitude recorded from neurons expressing mKate2 and SthK-bP when stimulated with 1 mW mm −2 470 nm light. Shown are individual data points, median and 25–75% interquartile range, n = 5; median peak current SthK-bP 1.37 nA. (F) Kinetics of SthK-bP; data obtained from traces when the stimulation intensity was 1 mW mm −2 (470 nm, 50 ms); n = 5. (G) Whole-cell responses to current injections from −400 to 700 pA in SthK-bP expressing hippocampal neuron. (H) Action potentials generated by repeated somatic current injection (1,000 pA, 600 ms, ISI 5 s) were blocked for 3 min by 3 470 nm light flashes at 40 second intervals (50 ms at 1 mW mm −2 ); same neuron as in (G) .
    Figure Legend Snippet: Characterization of a light-gated potassium channel in hippocampal neurons. (A) Maximum intensity projection of confocal images of hippocampal neurons 3 days after electroporation with DNA encoding SthK-bP and mKate2 (left: excitation 559 nm; right: excitation 488 nm). Lower images are close up of the regions indicated by dark boxes. (B) Single plane of the YFP signal of the region indicated in (A) (dark yellow box). (C) Left: Sample photocurrents evoked by a 50 ms light pulse (470 nm) of different intensity in a hippocampal neuron expressing mKate2 and SthK-bP. The holding potential was −70 mV. Right: Light intensity-response relationship fitted with a quadratic equation. Photocurrents are normalized to the maximum current recorded in each neuron. n = 5. (D) Left: Sample traces of photocurrents recorded from a SthK-bP expressing neuron when stimulated with 0.1 mW/mm 2 , 470 nm light (50 ms) while holding the cell at membrane potentials from −70 mV down to −115 mV. Baselines are aligned. Right: Current-voltage plot. A non-linear fit was applied to determine the K + equilibrium potential (−100.5 mV). (E) Photocurrent amplitude recorded from neurons expressing mKate2 and SthK-bP when stimulated with 1 mW mm −2 470 nm light. Shown are individual data points, median and 25–75% interquartile range, n = 5; median peak current SthK-bP 1.37 nA. (F) Kinetics of SthK-bP; data obtained from traces when the stimulation intensity was 1 mW mm −2 (470 nm, 50 ms); n = 5. (G) Whole-cell responses to current injections from −400 to 700 pA in SthK-bP expressing hippocampal neuron. (H) Action potentials generated by repeated somatic current injection (1,000 pA, 600 ms, ISI 5 s) were blocked for 3 min by 3 470 nm light flashes at 40 second intervals (50 ms at 1 mW mm −2 ); same neuron as in (G) .

    Techniques Used: Electroporation, Mass Spectrometry, Expressing, Generated, Injection

    Related Articles

    Synthesized:

    Article Title: Loss of native α-synuclein multimerization by strategically mutating its amphipathic helix causes abnormal vesicle interactions in neuronal cells
    Article Snippet: Wt mice (C57BL/6; 12 wk old) were from Charles River, Wilmington, MA. .. Plasmids pcDNA4/αS , pcDNA4/αS-3K , pcDNA4/αS-KLK , pcDNA4/αS-wt::YFP, pcDNA4/αS-3K::YFP , pcDNA4/αS-KLK::YFP , pcDNA4/αS-EIV::YFP ( ) and pCAX/dsRed ( ) have been described. αS-KLKEGR was synthesized as a GeneArt String DNA fragment (GeneArt/Life Technologies) and inserted into pcDNA4/TO/myc-His A (pcDNA4) with the In-Fusion HD Cloning Kit (Clontech). .. Plasmids mCherry-Rab5a-7 (Addgene plasmid # 55126), mCherry-Rab5a-7 (Addgene plasmid # 55127) mCherry-Rab11a-7 (Addgene plasmid # 55124), mCherry-SiT-N-15 (Addgene plasmid # 55133), mCherry-TFR-20 (Addgene plasmid # 55144), mCherry-ER-3 (Calreticulin; Addgene plasmid # 55041), mCherry-Endo-14 (RHOB; Addgene plasmid # 55040), mCherry-mito-7 (COX8A; Addgene plasmid # 55102), and mCherry-lysosomes-20 (LAMP1; Addgene plasmid # 55073) were kind gifts from Michael Davidson. pLVX-EF1α-IRES-mCherry vector was purchased from Clontech and viral constructs pLVX-IRES-mCherry/αS-wt, pLVX-IRES-mCherry/αS-3K and pLVX-IRES-mCherry/αS-KLK were generated by ligating SpeI/NotI digested PCR product into respective sites of pLVX-EF1α-IRES-mCherry.

    Article Title: A model of the onset of the senescence associated secretory phenotype after DNA damage induced senescence
    Article Snippet: In the first construct, the aim was to insert the T2A-mRuby2 sequence before the stop codon of Cre recombinase and in the second construct, the aim was to replace the Cre ORF with mRuby2 ORF. .. In brief, synthetic DNA fragments were synthesized either as gBlock (IDT) or as GeneArt string (Thermo Scientific). .. Four DNA fragments were synthesized, the first one contained 5’ 50 bp homology regions to the vector (targeting 50 nucleotide upstream of Cre ORF stop codon), chloramphenicol and ccdB cassettes and 3’ terminal 50 bp homology regions to the vector (targeting 50 nucleotide downstream of last amino acid coding codon of Cre ORF, i.e., condon preceding the Cre ORF stop codon).

    Article Title: OsMYC2, an essential factor for JA-inductive sakuranetin production in rice, interacts with MYC2-like proteins that enhance its transactivation ability
    Article Snippet: Amplified DNA fragments were self-ligated using the In-Fusion system (TaKaRa Bio USA, CA, USA) to obtain pEU-E01-His-Bls-GW-STOP. .. pEU-E01-6His-Bls-optOsMYC2 and pEU-E01-FLAG-optOsMYC2 The optimized CDS of OsMYC2 was synthesized using the GeneArt® Strings™ DNA Fragments service (Invitrogen, CA, USA), PCR-amplified in twice, and cloned into the pENTR/D-TOPO vector. .. The CDS was then inserted into the pEU-E01-6His-Bls-GW-STOP or pEU-E01-FLAG-GW-STOP vector by using the LR reaction.

    Article Title: Synthetic Light-Activated Ion Channels for Optogenetic Activation and Inhibition
    Article Snippet: The bPAC sequence is as previously published (Stierl et al., ). .. The SthK channel DNA sequence was synthesized by GeneArt Strings DNA Fragments (Life technologies, Thermo Fisher Scientific) according to the published amino acid sequence (Brams et al., ; Kesters et al., ) with codon usage optimized for Mus musculus . .. The DNA fragments were ligated and inserted into the oocyte expression vector pGEM-HE within N-terminal BamHI and C-terminal HindIII restriction sites.

    Clone Assay:

    Article Title: Loss of native α-synuclein multimerization by strategically mutating its amphipathic helix causes abnormal vesicle interactions in neuronal cells
    Article Snippet: Wt mice (C57BL/6; 12 wk old) were from Charles River, Wilmington, MA. .. Plasmids pcDNA4/αS , pcDNA4/αS-3K , pcDNA4/αS-KLK , pcDNA4/αS-wt::YFP, pcDNA4/αS-3K::YFP , pcDNA4/αS-KLK::YFP , pcDNA4/αS-EIV::YFP ( ) and pCAX/dsRed ( ) have been described. αS-KLKEGR was synthesized as a GeneArt String DNA fragment (GeneArt/Life Technologies) and inserted into pcDNA4/TO/myc-His A (pcDNA4) with the In-Fusion HD Cloning Kit (Clontech). .. Plasmids mCherry-Rab5a-7 (Addgene plasmid # 55126), mCherry-Rab5a-7 (Addgene plasmid # 55127) mCherry-Rab11a-7 (Addgene plasmid # 55124), mCherry-SiT-N-15 (Addgene plasmid # 55133), mCherry-TFR-20 (Addgene plasmid # 55144), mCherry-ER-3 (Calreticulin; Addgene plasmid # 55041), mCherry-Endo-14 (RHOB; Addgene plasmid # 55040), mCherry-mito-7 (COX8A; Addgene plasmid # 55102), and mCherry-lysosomes-20 (LAMP1; Addgene plasmid # 55073) were kind gifts from Michael Davidson. pLVX-EF1α-IRES-mCherry vector was purchased from Clontech and viral constructs pLVX-IRES-mCherry/αS-wt, pLVX-IRES-mCherry/αS-3K and pLVX-IRES-mCherry/αS-KLK were generated by ligating SpeI/NotI digested PCR product into respective sites of pLVX-EF1α-IRES-mCherry.

    Article Title: Genome-wide Specificity of Highly Efficient TALENs and CRISPR/Cas9 for T Cell Receptor Modification
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    Mutagenesis:

    Article Title: Genetic dissection of Escherichia coli's master diguanylate cyclase DgcE: Role of the N-terminal MASE1 domain and direct signal input from a GTPase partner system
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    Sequencing:

    Article Title: Genetic dissection of Escherichia coli's master diguanylate cyclase DgcE: Role of the N-terminal MASE1 domain and direct signal input from a GTPase partner system
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    Article Title: Synthetic Light-Activated Ion Channels for Optogenetic Activation and Inhibition
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    Recombinant:

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    Plasmid Preparation:

    Article Title: An efficient method for the preparation of preferentially heterodimerized recombinant S100A8/A9 coexpressed in Escherichia coli
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    Article Title: Genome-wide Specificity of Highly Efficient TALENs and CRISPR/Cas9 for T Cell Receptor Modification
    Article Snippet: .. Donor Construction and Targeted Integration PCR For the 200 bp α chain donor construct (TA200G), we ordered a GeneArt Strings DNA Fragment (Thermo Fisher Scientific) containing two 200 bp regions flanking the αT4 cutting site in the TRAC locus and restriction sites AsiSI and SbfI for cloning into a lentiviral transfer vector in antisense direction. .. A GFP-expression cassette comprising a phosphoglycerate kinase (PGK) promoter and a polyA signal was cloned between the two homology sites in antisense direction using MfeI and NheI.

    Article Title: OsMYC2, an essential factor for JA-inductive sakuranetin production in rice, interacts with MYC2-like proteins that enhance its transactivation ability
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    Expressing:

    Article Title: An efficient method for the preparation of preferentially heterodimerized recombinant S100A8/A9 coexpressed in Escherichia coli
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    Produced:

    Article Title: High-Yield Production of the Major Birch Pollen Allergen Bet v 1 With Allergen Immunogenicity in Nicotiana benthamiana
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    Amplification:

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    Polymerase Chain Reaction:

    Article Title: Genome-wide Specificity of Highly Efficient TALENs and CRISPR/Cas9 for T Cell Receptor Modification
    Article Snippet: .. Donor Construction and Targeted Integration PCR For the 200 bp α chain donor construct (TA200G), we ordered a GeneArt Strings DNA Fragment (Thermo Fisher Scientific) containing two 200 bp regions flanking the αT4 cutting site in the TRAC locus and restriction sites AsiSI and SbfI for cloning into a lentiviral transfer vector in antisense direction. .. A GFP-expression cassette comprising a phosphoglycerate kinase (PGK) promoter and a polyA signal was cloned between the two homology sites in antisense direction using MfeI and NheI.

    Article Title: OsMYC2, an essential factor for JA-inductive sakuranetin production in rice, interacts with MYC2-like proteins that enhance its transactivation ability
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    Construct:

    Article Title: Genome-wide Specificity of Highly Efficient TALENs and CRISPR/Cas9 for T Cell Receptor Modification
    Article Snippet: .. Donor Construction and Targeted Integration PCR For the 200 bp α chain donor construct (TA200G), we ordered a GeneArt Strings DNA Fragment (Thermo Fisher Scientific) containing two 200 bp regions flanking the αT4 cutting site in the TRAC locus and restriction sites AsiSI and SbfI for cloning into a lentiviral transfer vector in antisense direction. .. A GFP-expression cassette comprising a phosphoglycerate kinase (PGK) promoter and a polyA signal was cloned between the two homology sites in antisense direction using MfeI and NheI.

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    Thermo Fisher dna fragments
    Recombination of the gene drive cassette into the wildtype genome. Fibroblasts were coinfected with Towne-eGFP (WT) and GD-mCherry, and supernatant was used to infect fresh cells. a Representative examples of fluorescent viral plaques spreading on fibroblasts, expressing either eGFP only (left), mCherry only (middle), or both (right). Scale bar: 100 μm. b <t>PCR</t> for mCherry (upper band) and eGFP (lower band) on 48 recombinant viral genomes from two independent experiments. c Same as ( b ), after coinfection with GD-mCherry and a mutated Towne-eGFP (two biological replicates, 30 genomes). d PCR of homology arms and Sanger sequencing of 17 eGFP-mCherry expressing viral clones. Blue dots: single nucleotide polymorphisms (SNPs) from Towne strain; Red dots: TB40/E strain. e Fraction of SNPs of Towne or TB40/E origin alongside the hCMV genome. Each dot represents an individual SNP. Data combine two biological replicates after Oxford Nanopore sequencing. Coverage gives the number of reads, allowing multiple mapping. f Reconstruction of recombination history of individual hCMV genomes from long ( > 200 kb) Nanopore reads. One genome corresponds to one sequencing read and colors indicate the strain of origin. Gaps represent regions deleted compared to the reference genome, dashed lines indicate uncovered region. UL/US: unique long/short genome segments. GeneRuler 1 kb <t>DNA</t> ladder (ThermoFisher) was used in agarose gels as a size marker. The ladder is shown in the gel images, with the stronger band corresponding to 500 bp. Source data are provided as a Source Data file.
    Dna Fragments, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Recombination of the gene drive cassette into the wildtype genome. Fibroblasts were coinfected with Towne-eGFP (WT) and GD-mCherry, and supernatant was used to infect fresh cells. a Representative examples of fluorescent viral plaques spreading on fibroblasts, expressing either eGFP only (left), mCherry only (middle), or both (right). Scale bar: 100 μm. b PCR for mCherry (upper band) and eGFP (lower band) on 48 recombinant viral genomes from two independent experiments. c Same as ( b ), after coinfection with GD-mCherry and a mutated Towne-eGFP (two biological replicates, 30 genomes). d PCR of homology arms and Sanger sequencing of 17 eGFP-mCherry expressing viral clones. Blue dots: single nucleotide polymorphisms (SNPs) from Towne strain; Red dots: TB40/E strain. e Fraction of SNPs of Towne or TB40/E origin alongside the hCMV genome. Each dot represents an individual SNP. Data combine two biological replicates after Oxford Nanopore sequencing. Coverage gives the number of reads, allowing multiple mapping. f Reconstruction of recombination history of individual hCMV genomes from long ( > 200 kb) Nanopore reads. One genome corresponds to one sequencing read and colors indicate the strain of origin. Gaps represent regions deleted compared to the reference genome, dashed lines indicate uncovered region. UL/US: unique long/short genome segments. GeneRuler 1 kb DNA ladder (ThermoFisher) was used in agarose gels as a size marker. The ladder is shown in the gel images, with the stronger band corresponding to 500 bp. Source data are provided as a Source Data file.

    Journal: Nature Communications

    Article Title: Viral gene drive in herpesviruses

    doi: 10.1038/s41467-020-18678-0

    Figure Lengend Snippet: Recombination of the gene drive cassette into the wildtype genome. Fibroblasts were coinfected with Towne-eGFP (WT) and GD-mCherry, and supernatant was used to infect fresh cells. a Representative examples of fluorescent viral plaques spreading on fibroblasts, expressing either eGFP only (left), mCherry only (middle), or both (right). Scale bar: 100 μm. b PCR for mCherry (upper band) and eGFP (lower band) on 48 recombinant viral genomes from two independent experiments. c Same as ( b ), after coinfection with GD-mCherry and a mutated Towne-eGFP (two biological replicates, 30 genomes). d PCR of homology arms and Sanger sequencing of 17 eGFP-mCherry expressing viral clones. Blue dots: single nucleotide polymorphisms (SNPs) from Towne strain; Red dots: TB40/E strain. e Fraction of SNPs of Towne or TB40/E origin alongside the hCMV genome. Each dot represents an individual SNP. Data combine two biological replicates after Oxford Nanopore sequencing. Coverage gives the number of reads, allowing multiple mapping. f Reconstruction of recombination history of individual hCMV genomes from long ( > 200 kb) Nanopore reads. One genome corresponds to one sequencing read and colors indicate the strain of origin. Gaps represent regions deleted compared to the reference genome, dashed lines indicate uncovered region. UL/US: unique long/short genome segments. GeneRuler 1 kb DNA ladder (ThermoFisher) was used in agarose gels as a size marker. The ladder is shown in the gel images, with the stronger band corresponding to 500 bp. Source data are provided as a Source Data file.

    Article Snippet: All modifications were carried out by Gibson cloning (NEB, Ipswich, MA, USA), using PCR products from other plasmids or synthesized DNA fragments (GeneArt™ String™ fragments, ThermoFisher, USA).

    Techniques: Expressing, Polymerase Chain Reaction, Recombinant, Sequencing, Clone Assay, Nanopore Sequencing, Marker

    Schema for construction of S100A8/A9 coexpression vector and their protein expression in E. coli. (A) Expression vector for S100A8 or S100A9 were reconstructed into coexpression vectors by PCR amplification of each open reading frame and then subcloned by recombinase reaction. (B) Recombinant protein expression by each plasmid DNA for pET21-S100A8 (Lane 1), pET21-S100A9 (Lane2), pET21-S100A8-S100A9 (Lane 3), and pET21-S100A9-S100A8 (Lane 4) were confirmed by SDS-PAGE. Gels were stained with Coomassie Brilliant Blue (CBB).

    Journal: Biochemistry and Biophysics Reports

    Article Title: An efficient method for the preparation of preferentially heterodimerized recombinant S100A8/A9 coexpressed in Escherichia coli

    doi: 10.1016/j.bbrep.2016.03.009

    Figure Lengend Snippet: Schema for construction of S100A8/A9 coexpression vector and their protein expression in E. coli. (A) Expression vector for S100A8 or S100A9 were reconstructed into coexpression vectors by PCR amplification of each open reading frame and then subcloned by recombinase reaction. (B) Recombinant protein expression by each plasmid DNA for pET21-S100A8 (Lane 1), pET21-S100A9 (Lane2), pET21-S100A8-S100A9 (Lane 3), and pET21-S100A9-S100A8 (Lane 4) were confirmed by SDS-PAGE. Gels were stained with Coomassie Brilliant Blue (CBB).

    Article Snippet: 2.1 Construction of recombinant S100A8/A9 plasmid DNA The gene fragments encoding human S100A8 (Uniprot: P05109 ) and S100A9 (Uniprot: P06702 ) were prepared by GeneArt Strings DNA fragment gene synthesis service (Life Technologies) with the sequence optimized for E. coli protein expression.

    Techniques: Plasmid Preparation, Expressing, Polymerase Chain Reaction, Amplification, Recombinant, SDS Page, Staining

    Transcriptional activities of OsMYC2, OsMYL1, and OsMYL2. ( a ) Schematic representation of the vectors used for the transient expression assays. GAL4DBD: GAL4 DNA-binding domain, FLUC: firefly luciferase, RLUC: renilla luciferase, Pro35S, CaMV 35 S promoter, ProUbi: maize ubiquitin promoter, NosT: nopaline synthase terminator. ( b ) Relative luciferase activities in the bombarded rice leaves. Data are presented as FLUC/RLUC ± standard error. n = 6. *P

    Journal: Scientific Reports

    Article Title: OsMYC2, an essential factor for JA-inductive sakuranetin production in rice, interacts with MYC2-like proteins that enhance its transactivation ability

    doi: 10.1038/srep40175

    Figure Lengend Snippet: Transcriptional activities of OsMYC2, OsMYL1, and OsMYL2. ( a ) Schematic representation of the vectors used for the transient expression assays. GAL4DBD: GAL4 DNA-binding domain, FLUC: firefly luciferase, RLUC: renilla luciferase, Pro35S, CaMV 35 S promoter, ProUbi: maize ubiquitin promoter, NosT: nopaline synthase terminator. ( b ) Relative luciferase activities in the bombarded rice leaves. Data are presented as FLUC/RLUC ± standard error. n = 6. *P

    Article Snippet: pEU-E01-6His-Bls-optOsMYC2 and pEU-E01-FLAG-optOsMYC2 The optimized CDS of OsMYC2 was synthesized using the GeneArt® Strings™ DNA Fragments service (Invitrogen, CA, USA), PCR-amplified in twice, and cloned into the pENTR/D-TOPO vector.

    Techniques: Expressing, Binding Assay, Luciferase

    Characterization of a light-gated potassium channel in hippocampal neurons. (A) Maximum intensity projection of confocal images of hippocampal neurons 3 days after electroporation with DNA encoding SthK-bP and mKate2 (left: excitation 559 nm; right: excitation 488 nm). Lower images are close up of the regions indicated by dark boxes. (B) Single plane of the YFP signal of the region indicated in (A) (dark yellow box). (C) Left: Sample photocurrents evoked by a 50 ms light pulse (470 nm) of different intensity in a hippocampal neuron expressing mKate2 and SthK-bP. The holding potential was −70 mV. Right: Light intensity-response relationship fitted with a quadratic equation. Photocurrents are normalized to the maximum current recorded in each neuron. n = 5. (D) Left: Sample traces of photocurrents recorded from a SthK-bP expressing neuron when stimulated with 0.1 mW/mm 2 , 470 nm light (50 ms) while holding the cell at membrane potentials from −70 mV down to −115 mV. Baselines are aligned. Right: Current-voltage plot. A non-linear fit was applied to determine the K + equilibrium potential (−100.5 mV). (E) Photocurrent amplitude recorded from neurons expressing mKate2 and SthK-bP when stimulated with 1 mW mm −2 470 nm light. Shown are individual data points, median and 25–75% interquartile range, n = 5; median peak current SthK-bP 1.37 nA. (F) Kinetics of SthK-bP; data obtained from traces when the stimulation intensity was 1 mW mm −2 (470 nm, 50 ms); n = 5. (G) Whole-cell responses to current injections from −400 to 700 pA in SthK-bP expressing hippocampal neuron. (H) Action potentials generated by repeated somatic current injection (1,000 pA, 600 ms, ISI 5 s) were blocked for 3 min by 3 470 nm light flashes at 40 second intervals (50 ms at 1 mW mm −2 ); same neuron as in (G) .

    Journal: Frontiers in Neuroscience

    Article Title: Synthetic Light-Activated Ion Channels for Optogenetic Activation and Inhibition

    doi: 10.3389/fnins.2018.00643

    Figure Lengend Snippet: Characterization of a light-gated potassium channel in hippocampal neurons. (A) Maximum intensity projection of confocal images of hippocampal neurons 3 days after electroporation with DNA encoding SthK-bP and mKate2 (left: excitation 559 nm; right: excitation 488 nm). Lower images are close up of the regions indicated by dark boxes. (B) Single plane of the YFP signal of the region indicated in (A) (dark yellow box). (C) Left: Sample photocurrents evoked by a 50 ms light pulse (470 nm) of different intensity in a hippocampal neuron expressing mKate2 and SthK-bP. The holding potential was −70 mV. Right: Light intensity-response relationship fitted with a quadratic equation. Photocurrents are normalized to the maximum current recorded in each neuron. n = 5. (D) Left: Sample traces of photocurrents recorded from a SthK-bP expressing neuron when stimulated with 0.1 mW/mm 2 , 470 nm light (50 ms) while holding the cell at membrane potentials from −70 mV down to −115 mV. Baselines are aligned. Right: Current-voltage plot. A non-linear fit was applied to determine the K + equilibrium potential (−100.5 mV). (E) Photocurrent amplitude recorded from neurons expressing mKate2 and SthK-bP when stimulated with 1 mW mm −2 470 nm light. Shown are individual data points, median and 25–75% interquartile range, n = 5; median peak current SthK-bP 1.37 nA. (F) Kinetics of SthK-bP; data obtained from traces when the stimulation intensity was 1 mW mm −2 (470 nm, 50 ms); n = 5. (G) Whole-cell responses to current injections from −400 to 700 pA in SthK-bP expressing hippocampal neuron. (H) Action potentials generated by repeated somatic current injection (1,000 pA, 600 ms, ISI 5 s) were blocked for 3 min by 3 470 nm light flashes at 40 second intervals (50 ms at 1 mW mm −2 ); same neuron as in (G) .

    Article Snippet: The SthK channel DNA sequence was synthesized by GeneArt Strings DNA Fragments (Life technologies, Thermo Fisher Scientific) according to the published amino acid sequence (Brams et al., ; Kesters et al., ) with codon usage optimized for Mus musculus .

    Techniques: Electroporation, Mass Spectrometry, Expressing, Generated, Injection