Journal: Cell chemical biology

Article Title: A Strategic Approach for Fluorescence Imaging of Membrane Proteins in a Native-Like Environment

doi: 10.1016/j.chembiol.2019.11.008

Figure Lengend Snippet: (A) Expression yield of PglC (left) or PglA (right) variants normalized to the expression of the corresponding WT protein, in the presence or absence of BCN. Expression was quantified by anti-His western blot. (B) Heatmap representing the amber suppression efficiency at individual sites on PglC (left) and PglA (right) (see Figure S2 for details on the structure prediction). The structure of PglC from C. jejuni was predicted with the online server iTasser using the PglC crystal structure (PDB: 5W7L) from C. concisus as template (Ray et al., 2018; Roy et al., 2010; Yang et al., 2015; Zhang, 2008). The position of PglC with respect to the membrane plane was determined using the server “Orientations of Proteins in Membranes” Lomize et al., 2012). Positions selected for the insertions of non-canonical amino acids were selected based on their position within this predicted structure (surfaced exposed and in a diverse physico-chemical environment) and also based on the alignment of 15,000 sequences (non-conserved residues) Lukose et al., 2015). The structure of PglA from C. jejuni was predicted with the online server iTasser using the structure of the glycosyl transferases WbnH (PDB: 4XYW) from E. coli as the template (Martinez-Fleites et al., 2006; Roy et al., 2010; Yang et al., 2015; Zhang, 2008). Position of PglA with respect to the membrane plane was also determined using the server “Orientations of Proteins in Membranes”. Amber suppression efficiencies were calculated as the difference between the quantity of variant expressed in the presence and absence of BCN, normalized by the quantity of expressed WT protein. See also Tables S2 and S3, and Figure S2.

Article Snippet: Escherichia coli C321.ΔprfA (GRO) , Lajoie et al., 2013 , Addgene bacterial strain #48998.

Techniques: Expressing, Western Blot, Membrane, Variant Assay

Journal: Cell chemical biology

Article Title: A Strategic Approach for Fluorescence Imaging of Membrane Proteins in a Native-Like Environment

doi: 10.1016/j.chembiol.2019.11.008

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Escherichia coli C321.ΔprfA (GRO) , Lajoie et al., 2013 , Addgene bacterial strain #48998.

Techniques: Recombinant, Plasmid Preparation, Mutagenesis, Sequencing, Expressing, Software

Journal: Synthetic and Systems Biotechnology

Article Title: A linear DNA template-based framework for site-specific unnatural amino acid incorporation

doi: 10.1016/j.synbio.2021.07.003

Figure Lengend Snippet: Screening of 14 cell extracts. In this experiment, pPaF was selected for the UNAA embedding screening of 14 kinds of cell extracts. A total of 14 extracts were selected, including K-12 series (K-12, K-12 ΔtnaA, K-12 ΔtnaAΔsdaB), commercial series (BL21 (DE3), BL21 ΔserB, Rosetta (DE3), Rosetta-gami B (DE3), Origami, Origami B), rEc series (rEc. 13, rEc. 13. ΔA), EcAR7 series (EcAR7 ΔA ΔSer), C321 series (C321, C321. ΔA).

Article Snippet: ΔA [ ] (Addgene #69495)), EcAR7 series (EcAR7 ΔA ΔSer [ ]) (Addgene #52055), C321 series (C321 [ ] (Addgene #48999), C321.

Techniques: