Journal: Frontiers in Plant Science

Article Title: Germacrene A synthase in yarrow (Achillea millefolium ) is an enzyme with mixed substrate specificity: gene cloning, functional characterization and expression analysis

doi: 10.3389/fpls.2015.00111

Figure Lengend Snippet: Heterologus expression of AmGAS . (A) sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), the numbers indicate—1: protein marker, 2: induced recombinant strain containing pET-26b (+) plus AmGAS gene after 6 h, 3, after 4 h; 4, after 2 h; 5, non-induced and 6, E. coli BL21 (DE3) without plasmid, and (B) western blotting, the numbers denote—1, non-induced recombinant strain containing pET-26b (+) plus AmGAS gene; 2, induced protein after 2 h; 3, after 4 h; 4, after 6 h and 5, protein marker. Arrows show the band that belongs to the recombinant protein of AmGAS.

Article Snippet: The AmGAS PCR product and pET-26b (+) expression vector were digested with NcoI and XhoI restriction enzymes according to the manufacturer's protocol (New England Biolabs, Ipswich, MA, USA).

Techniques: Expressing, Polyacrylamide Gel Electrophoresis, SDS Page, Marker, Recombinant, Plasmid Preparation, Western Blot

Journal: Microbial Cell Factories

Article Title: Secretion of the human parathyroid hormone through a microcin type I secretion system in Escherichia coli

doi: 10.1186/s12934-024-02552-5

Figure Lengend Snippet: Sequences containing the gene fusions cloned into pET-26b(+). Nucleotide sequences of the inserts and of the vector adjacencies are indicated. In grey and boxed, restriction sites employed for cloning the fusions. Start and stop codons are in bold and Shine-Dalgarno sequences are underlined. The amino acid sequence of the fusion products is indicated. Sequences corresponding to SD V and PTH variants are in blue and red, respectively. ( A ) sd v -pth34-6xhis (229 bp). ( B ) sd v -pth84-6xhis (374 bp)

Article Snippet: The gene fusions sd v -pth34 and sd v -pth84 were provided by GenScript cloned into the pET-26b(+) vector.

Techniques: Clone Assay, Plasmid Preparation, Cloning, Sequencing

Journal: Microbial Cell Factories

Article Title: Secretion of the human parathyroid hormone through a microcin type I secretion system in Escherichia coli

doi: 10.1186/s12934-024-02552-5

Figure Lengend Snippet: Secretion of PTH peptides: supernatants. ( A ) Tricine SDS-PAGE of supernatants from cultures of control and experimental strains. Aliquots of 10 µl of each supernatant were loaded. (1): 1 µl of PTH84 (1 µg); (2): BL21(DE3) (pExAB; pET-PTH84); (3): BL21(DE3) (pET-PTH84); (4): protein ladder (5 µl); (5): BL21(DE3) (pExAB); (6): BL21(DE3) (pExAB; pET-PTH34); (7): BL21(DE3) (pET-PTH34); (8): 1 µl of PTH34 (1 µg). Asterisks, bands analysed by mass spectrometry. ( B ) Mass spectrometry analyses from bands marked in A. PTH84-6xHis and PTH34-6xHis sequences were included in the data base of the analysis. Red, peptides detected by mass spectrometry

Article Snippet: The gene fusions sd v -pth34 and sd v -pth84 were provided by GenScript cloned into the pET-26b(+) vector.

Techniques: SDS Page, Control, Mass Spectrometry

Journal: Microbial Cell Factories

Article Title: Secretion of the human parathyroid hormone through a microcin type I secretion system in Escherichia coli

doi: 10.1186/s12934-024-02552-5

Figure Lengend Snippet: Intracellular PTH peptides: cell lysates. ( A ) Tricine SDS-PAGE of BL21(DE3) (pExAB; pET-PTH84) lysate. Lines: 1, 1 µl of PTH84 (1 µg); 2, 10 µl of cell lysate. (1) and (2), bands analysed by mass spectrometry. ( B ) Mass spectrometry analyses. Data base for band (1) included the unprocessed SD V -PTH84-6xHis sequence, and for band (2), the processed PTH84-6xHis sequence. Red, peptides detected by mass spectrometry. Arrow, specific cleavage site performed by the MccV T1SS

Article Snippet: The gene fusions sd v -pth34 and sd v -pth84 were provided by GenScript cloned into the pET-26b(+) vector.

Techniques: SDS Page, Mass Spectrometry, Sequencing

Journal: Microbial Cell Factories

Article Title: Secretion of the human parathyroid hormone through a microcin type I secretion system in Escherichia coli

doi: 10.1186/s12934-024-02552-5

Figure Lengend Snippet: Western blot assay for the identification of PTH84-6xHis peptide forms. Membrane transferred from Tricine SDS-PAGE loaded with samples from BL21(DE3) (pExAB; pET-PTH84) culture. Detection with Anti-His tag antibody. (1): protein ladder (5 µl); (2): 15 µl of supernatant; (3): 15 µl of cell lysate

Article Snippet: The gene fusions sd v -pth34 and sd v -pth84 were provided by GenScript cloned into the pET-26b(+) vector.

Techniques: Western Blot, Membrane, SDS Page