Structured Review

5 PRIME gfp control vector
Western Blot of WT <t>GFP</t> and GFP Y39TAG mutant expression in a cell-free translation system. Synthesis of WT GFP and the GFP Y39TAG mutant was performed using the <t>RTS</t> E. coli HY Kit, to which the corresponding plasmid (500 µg/mL), purified Mj TyrRS and cognate suppressor Mj tRNA CUA (tRNA) or T-stem modified tRNA CUA Opt (denoted as *) were added. (A) Expression of WT GFP and the GFP Y39TAG mutant in the presence of Mj TyrRS (300 µg/mL) and synthetic Mj tRNA CUA (60 µg/mL). The band at 28 kDa corresponds to full-length GFP. (B) Western blot analysis demonstrates enhanced GFP Y39TAG protein expression as a function of increased Mj TyrRS concentrations in a cell-free reaction medium supplied with Mj tRNA CUA (60 µg/mL – top panel and 450 µg/mL – bottom panel). ( C ) Dependence of GFP Y39TAG yield on the type and concentration of nonsense suppressor, as visualized by Western blot.
Gfp Control Vector, supplied by 5 PRIME, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/gfp control vector/product/5 PRIME
Average 88 stars, based on 1 article reviews
Price from $9.99 to $1999.99
gfp control vector - by Bioz Stars, 2020-08
88/100 stars

Images

1) Product Images from "Enhanced Yield of Recombinant Proteins with Site-Specifically Incorporated Unnatural Amino Acids Using a Cell-Free Expression System"

Article Title: Enhanced Yield of Recombinant Proteins with Site-Specifically Incorporated Unnatural Amino Acids Using a Cell-Free Expression System

Journal: PLoS ONE

doi: 10.1371/journal.pone.0068363

Western Blot of WT GFP and GFP Y39TAG mutant expression in a cell-free translation system. Synthesis of WT GFP and the GFP Y39TAG mutant was performed using the RTS E. coli HY Kit, to which the corresponding plasmid (500 µg/mL), purified Mj TyrRS and cognate suppressor Mj tRNA CUA (tRNA) or T-stem modified tRNA CUA Opt (denoted as *) were added. (A) Expression of WT GFP and the GFP Y39TAG mutant in the presence of Mj TyrRS (300 µg/mL) and synthetic Mj tRNA CUA (60 µg/mL). The band at 28 kDa corresponds to full-length GFP. (B) Western blot analysis demonstrates enhanced GFP Y39TAG protein expression as a function of increased Mj TyrRS concentrations in a cell-free reaction medium supplied with Mj tRNA CUA (60 µg/mL – top panel and 450 µg/mL – bottom panel). ( C ) Dependence of GFP Y39TAG yield on the type and concentration of nonsense suppressor, as visualized by Western blot.
Figure Legend Snippet: Western Blot of WT GFP and GFP Y39TAG mutant expression in a cell-free translation system. Synthesis of WT GFP and the GFP Y39TAG mutant was performed using the RTS E. coli HY Kit, to which the corresponding plasmid (500 µg/mL), purified Mj TyrRS and cognate suppressor Mj tRNA CUA (tRNA) or T-stem modified tRNA CUA Opt (denoted as *) were added. (A) Expression of WT GFP and the GFP Y39TAG mutant in the presence of Mj TyrRS (300 µg/mL) and synthetic Mj tRNA CUA (60 µg/mL). The band at 28 kDa corresponds to full-length GFP. (B) Western blot analysis demonstrates enhanced GFP Y39TAG protein expression as a function of increased Mj TyrRS concentrations in a cell-free reaction medium supplied with Mj tRNA CUA (60 µg/mL – top panel and 450 µg/mL – bottom panel). ( C ) Dependence of GFP Y39TAG yield on the type and concentration of nonsense suppressor, as visualized by Western blot.

Techniques Used: Western Blot, Mutagenesis, Expressing, Plasmid Preparation, Purification, Modification, Concentration Assay

Related Articles

Plasmid Preparation:

Article Title: Enhanced Yield of Recombinant Proteins with Site-Specifically Incorporated Unnatural Amino Acids Using a Cell-Free Expression System
Article Snippet: .. GFP Mutants The X-ray crystallographic structure of template GFP (PDB accession number: 1EMA), encoded by the GFP control vector (RTS, 5 PRIME, Hamburg, Germany), was analyzed to choose sites for UAA incorporation. ..

Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 92
    5 PRIME gfp tagged trp47 constructs plasmids
    A MYND-binding domain is responsible for <t>TRP47</t> nuclear translocation. (A) Schematic showing <t>GFP-tagged</t> TRP47 expression constructs. Red boxes indicate presence of the MYND-binding domain (MBD). Blue boxes represent the seven 19-mer repeats of the TRP47 tandem repeat region. Open space between two lines indicates a deletion mutation. Subscripts on construct labels indicate which amino acid residues are included. Constructs shown are full-length (FL), His-tagged MBD (MBD-His), full-length TRP47 without the MBD (No Motif), N-terminal (N), truncated N-terminal without the MBD (Ntrunc), tandem repeat-C-terminal overlapping the MBD (TRC), C-terminal (C), K49R mutant (K49R), and K71R mutant (K71R). (B) Ectopically expressed GFP-tagged constructs fluoresce green and DAPI-stained nuclei fluoresce blue. The FL, N, TRC, His-MBD, K49R, and K71R constructs contained the MBD (marked by asterisks), while the Ntrunc, C, and No Motif constructs did not. The FL and His-MBD TRP47 constructs exhibited strong nuclear localization and the N, TRC, and K71R TRP47 constructs exhibited nuclear and diffuse cytoplasmic localization. Exclusively cytoplasmic localization was observed with the Ntrunc, C, No Motif, and K49R mutant TRP47 constructs. The GFP negative control construct was also observed only in the cytoplasm. Due to differences in expression levels of the constructs, different exposure times were used to image each construct.
    Gfp Tagged Trp47 Constructs Plasmids, supplied by 5 PRIME, used in various techniques. Bioz Stars score: 92/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/gfp tagged trp47 constructs plasmids/product/5 PRIME
    Average 92 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    gfp tagged trp47 constructs plasmids - by Bioz Stars, 2020-08
    92/100 stars
      Buy from Supplier

    88
    5 PRIME gfp control vector
    Western Blot of WT <t>GFP</t> and GFP Y39TAG mutant expression in a cell-free translation system. Synthesis of WT GFP and the GFP Y39TAG mutant was performed using the <t>RTS</t> E. coli HY Kit, to which the corresponding plasmid (500 µg/mL), purified Mj TyrRS and cognate suppressor Mj tRNA CUA (tRNA) or T-stem modified tRNA CUA Opt (denoted as *) were added. (A) Expression of WT GFP and the GFP Y39TAG mutant in the presence of Mj TyrRS (300 µg/mL) and synthetic Mj tRNA CUA (60 µg/mL). The band at 28 kDa corresponds to full-length GFP. (B) Western blot analysis demonstrates enhanced GFP Y39TAG protein expression as a function of increased Mj TyrRS concentrations in a cell-free reaction medium supplied with Mj tRNA CUA (60 µg/mL – top panel and 450 µg/mL – bottom panel). ( C ) Dependence of GFP Y39TAG yield on the type and concentration of nonsense suppressor, as visualized by Western blot.
    Gfp Control Vector, supplied by 5 PRIME, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/gfp control vector/product/5 PRIME
    Average 88 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    gfp control vector - by Bioz Stars, 2020-08
    88/100 stars
      Buy from Supplier

    93
    5 PRIME transfection
    Western Blot of WT <t>GFP</t> and GFP Y39TAG mutant expression in a cell-free translation system. Synthesis of WT GFP and the GFP Y39TAG mutant was performed using the <t>RTS</t> E. coli HY Kit, to which the corresponding plasmid (500 µg/mL), purified Mj TyrRS and cognate suppressor Mj tRNA CUA (tRNA) or T-stem modified tRNA CUA Opt (denoted as *) were added. (A) Expression of WT GFP and the GFP Y39TAG mutant in the presence of Mj TyrRS (300 µg/mL) and synthetic Mj tRNA CUA (60 µg/mL). The band at 28 kDa corresponds to full-length GFP. (B) Western blot analysis demonstrates enhanced GFP Y39TAG protein expression as a function of increased Mj TyrRS concentrations in a cell-free reaction medium supplied with Mj tRNA CUA (60 µg/mL – top panel and 450 µg/mL – bottom panel). ( C ) Dependence of GFP Y39TAG yield on the type and concentration of nonsense suppressor, as visualized by Western blot.
    Transfection, supplied by 5 PRIME, used in various techniques. Bioz Stars score: 93/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/transfection/product/5 PRIME
    Average 93 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    transfection - by Bioz Stars, 2020-08
    93/100 stars
      Buy from Supplier

    Image Search Results


    A MYND-binding domain is responsible for TRP47 nuclear translocation. (A) Schematic showing GFP-tagged TRP47 expression constructs. Red boxes indicate presence of the MYND-binding domain (MBD). Blue boxes represent the seven 19-mer repeats of the TRP47 tandem repeat region. Open space between two lines indicates a deletion mutation. Subscripts on construct labels indicate which amino acid residues are included. Constructs shown are full-length (FL), His-tagged MBD (MBD-His), full-length TRP47 without the MBD (No Motif), N-terminal (N), truncated N-terminal without the MBD (Ntrunc), tandem repeat-C-terminal overlapping the MBD (TRC), C-terminal (C), K49R mutant (K49R), and K71R mutant (K71R). (B) Ectopically expressed GFP-tagged constructs fluoresce green and DAPI-stained nuclei fluoresce blue. The FL, N, TRC, His-MBD, K49R, and K71R constructs contained the MBD (marked by asterisks), while the Ntrunc, C, and No Motif constructs did not. The FL and His-MBD TRP47 constructs exhibited strong nuclear localization and the N, TRC, and K71R TRP47 constructs exhibited nuclear and diffuse cytoplasmic localization. Exclusively cytoplasmic localization was observed with the Ntrunc, C, No Motif, and K49R mutant TRP47 constructs. The GFP negative control construct was also observed only in the cytoplasm. Due to differences in expression levels of the constructs, different exposure times were used to image each construct.

    Journal: PLoS ONE

    Article Title: Ehrlichia chaffeensis TRP47 enters the nucleus via a MYND-binding domain-dependent mechanism and predominantly binds enhancers of host genes associated with signal transduction, cytoskeletal organization, and immune response

    doi: 10.1371/journal.pone.0205983

    Figure Lengend Snippet: A MYND-binding domain is responsible for TRP47 nuclear translocation. (A) Schematic showing GFP-tagged TRP47 expression constructs. Red boxes indicate presence of the MYND-binding domain (MBD). Blue boxes represent the seven 19-mer repeats of the TRP47 tandem repeat region. Open space between two lines indicates a deletion mutation. Subscripts on construct labels indicate which amino acid residues are included. Constructs shown are full-length (FL), His-tagged MBD (MBD-His), full-length TRP47 without the MBD (No Motif), N-terminal (N), truncated N-terminal without the MBD (Ntrunc), tandem repeat-C-terminal overlapping the MBD (TRC), C-terminal (C), K49R mutant (K49R), and K71R mutant (K71R). (B) Ectopically expressed GFP-tagged constructs fluoresce green and DAPI-stained nuclei fluoresce blue. The FL, N, TRC, His-MBD, K49R, and K71R constructs contained the MBD (marked by asterisks), while the Ntrunc, C, and No Motif constructs did not. The FL and His-MBD TRP47 constructs exhibited strong nuclear localization and the N, TRC, and K71R TRP47 constructs exhibited nuclear and diffuse cytoplasmic localization. Exclusively cytoplasmic localization was observed with the Ntrunc, C, No Motif, and K49R mutant TRP47 constructs. The GFP negative control construct was also observed only in the cytoplasm. Due to differences in expression levels of the constructs, different exposure times were used to image each construct.

    Article Snippet: Ectopic expression of GFP-tagged TRP47 constructs Plasmids for transfection (pAcGFP1-C FL, N, Ntrunc, TRC, C, MBD, No Motif, K49R, K71R, GFP control) were isolated from 150 mL overnight E . coli cultures with the PerfectPrep EndoFree Plasmid Maxi Kit (5 PRIME) and quantitated using a spectrophotometer.

    Techniques: Binding Assay, Translocation Assay, Expressing, Construct, Mutagenesis, Staining, Negative Control

    Western Blot of WT GFP and GFP Y39TAG mutant expression in a cell-free translation system. Synthesis of WT GFP and the GFP Y39TAG mutant was performed using the RTS E. coli HY Kit, to which the corresponding plasmid (500 µg/mL), purified Mj TyrRS and cognate suppressor Mj tRNA CUA (tRNA) or T-stem modified tRNA CUA Opt (denoted as *) were added. (A) Expression of WT GFP and the GFP Y39TAG mutant in the presence of Mj TyrRS (300 µg/mL) and synthetic Mj tRNA CUA (60 µg/mL). The band at 28 kDa corresponds to full-length GFP. (B) Western blot analysis demonstrates enhanced GFP Y39TAG protein expression as a function of increased Mj TyrRS concentrations in a cell-free reaction medium supplied with Mj tRNA CUA (60 µg/mL – top panel and 450 µg/mL – bottom panel). ( C ) Dependence of GFP Y39TAG yield on the type and concentration of nonsense suppressor, as visualized by Western blot.

    Journal: PLoS ONE

    Article Title: Enhanced Yield of Recombinant Proteins with Site-Specifically Incorporated Unnatural Amino Acids Using a Cell-Free Expression System

    doi: 10.1371/journal.pone.0068363

    Figure Lengend Snippet: Western Blot of WT GFP and GFP Y39TAG mutant expression in a cell-free translation system. Synthesis of WT GFP and the GFP Y39TAG mutant was performed using the RTS E. coli HY Kit, to which the corresponding plasmid (500 µg/mL), purified Mj TyrRS and cognate suppressor Mj tRNA CUA (tRNA) or T-stem modified tRNA CUA Opt (denoted as *) were added. (A) Expression of WT GFP and the GFP Y39TAG mutant in the presence of Mj TyrRS (300 µg/mL) and synthetic Mj tRNA CUA (60 µg/mL). The band at 28 kDa corresponds to full-length GFP. (B) Western blot analysis demonstrates enhanced GFP Y39TAG protein expression as a function of increased Mj TyrRS concentrations in a cell-free reaction medium supplied with Mj tRNA CUA (60 µg/mL – top panel and 450 µg/mL – bottom panel). ( C ) Dependence of GFP Y39TAG yield on the type and concentration of nonsense suppressor, as visualized by Western blot.

    Article Snippet: GFP Mutants The X-ray crystallographic structure of template GFP (PDB accession number: 1EMA), encoded by the GFP control vector (RTS, 5 PRIME, Hamburg, Germany), was analyzed to choose sites for UAA incorporation.

    Techniques: Western Blot, Mutagenesis, Expressing, Plasmid Preparation, Purification, Modification, Concentration Assay