recombinant plasmid pet-28b-ftsz  (GenScript corporation)

Journal: Acta Crystallographica. Section F, Structural Biology Communications

Article Title: Crystal structure of the kringle domain of human receptor tyrosine kinase-like orphan receptor 1 (hROR1)

doi: 10.1107/S2053230X22003855

Figure Lengend Snippet: Macromolecule-production information

Article Snippet: The codon-optimized cDNA construct encoding human hROR1-KRD (UniProt entry Q01973, residues 312–393) was synthesized by Genewiz and subcloned into a modified pET-28b-SUMO recombinant expression plasmid (Novagen) using in-frame 5′-BamHI and 3′-NotI restriction sites, yielding a final construct bearing an N-terminal 8×His-SUMO fusion (Table 1 ) and cloning scars corresponding to the amino-acid sequences encoded by the restriction nucleases BamHI at the N-terminus (Gly-Ser) and NotI at the C-terminus (Ala-Ala-Ala); the latter precedes an in-frame stop codon already embedded in the expression vector.

Techniques: Expressing, Plasmid Preparation, Modification, Sequencing, Construct, Produced, Crystallization Assay

Journal: Frontiers in Plant Science

Article Title: DNA-Free Genome Editing of Brassica oleracea and B. rapa Protoplasts Using CRISPR-Cas9 Ribonucleoprotein Complexes

doi: 10.3389/fpls.2018.01594

Figure Lengend Snippet: Mutation rates based on deep sequencing of target regions in FRI and PDS genes and analysis with Cas-Analyzer.

Article Snippet: The pET28b-His-Cas9 vector containing recombinant Cas9 gene was obtained from Addgene (plasmid ID 47327) and expressed in E. coli Rosetta cells (Novagen).

Techniques: Mutagenesis, Sequencing

Journal: Frontiers in Plant Science

Article Title: DNA-Free Genome Editing of Brassica oleracea and B. rapa Protoplasts Using CRISPR-Cas9 Ribonucleoprotein Complexes

doi: 10.3389/fpls.2018.01594

Figure Lengend Snippet: Distribution of the most frequent alleles identified with CRISPResso around cleavage sites in B. rapa. Sequences were obtained after transfection with 60 μg Cas9 preassembled with 60 μg of sgRNA-FRI1, sgRNA-FRI4, sgRNA-PDS1, or sgRNA-PDS2. The horizontal line indicates the PAM region, the vertical dashed line indicates the predicted cleavage site. Mutations are shown in bold font (substitutions), highlighted with red rectangles (insertions), or marked with horizontal dashed lines (deletions).

Article Snippet: The pET28b-His-Cas9 vector containing recombinant Cas9 gene was obtained from Addgene (plasmid ID 47327) and expressed in E. coli Rosetta cells (Novagen).

Techniques: Transfection