Five high-abundance protein spots identified by MALDI-TOF/TOF.

a<p>The expression level of <i>P. putida</i> JUCT1 grown in nutrient medium without solvent.</p>b<p>The expression level of <i>P. putida</i> JUCT1 grown in the presence of 60% (v/v) cyclohexane.</p

The oxidation of cyclohexane by <i>E. coli</i> strains.

<p>Incubation condition: 37°C for 8 h with cyclohexane (1%, w/v).</p

Cell growth of recombinant <i>E. coli</i> JM109 strains in the presence of 4% (v/v) cyclohexane.

<p>The recombinant strains were cultured at 37°C and cyclohexane (CH) was added when OD₆₆₀ reached 0.2 (indicated by the arrow). □, pQE-80L (as the control); ▾, pQE-<i>mmsB</i>; •, pQE- <i>tsf</i>; △, pQE-<i>PSEEN0851</i>.</p

Effect of organic solvents on cell growth of <i>P. putida</i> JUCT1 (open bar) and its parent strain JUCS (gray bar).

<p>The strains were initially grown in nutrient medium at 37°C till OD₆₆₀ reached 0.2, and then 60% (v/v) organic solvent was added for further incubation of 5 h.</p

MALDI-TOF/TOF analysis of peptides from 5 high-abundance proteins.

a<p>Number of peptides identified for individual protein in MALDI-TOF/TOF analysis.</p>b<p>Number of unique peptides identified for individual protein.</p>c<p>Total amino acid sequence of peptides identified for individual protein.</p>d<p>Percentage of amino acid sequence covered by peptides of individual protein.</p

Colony formation of <i>E. coli</i> JM109 strains over-expressing <i>mmsB</i>, <i>tsf</i>, and <i>PSEEN0851</i> on LBGMg agar overlaid with decalin after 24 h incubation at 37°C. JM109 carrying empty pQE-80L was used as the control.

<p>Colony formation of <i>E. coli</i> JM109 strains over-expressing <i>mmsB</i>, <i>tsf</i>, and <i>PSEEN0851</i> on LBGMg agar overlaid with decalin after 24 h incubation at 37°C. JM109 carrying empty pQE-80L was used as the control.</p

SDS-PAGE analysis of expression of <i>mmsB</i>, <i>PSEEN0851</i>, and <i>tsf</i> in <i>E. coli</i> JM109.

<p>The transformants were grown in LB at 37°C and induced by 1 mM IPTG. Lanes: M, molecular mass marker; 1, pQE-80L (as control); 2, pQE-<i>mmsB</i>; 3, pQE-<i>PSEEN0851</i>; 4, pQE-<i>tsf</i>. Arrowheads indicate the locations of recombinant proteins.</p

Facile Preparation of Well-Defined Hydrophilic Core–Shell Upconversion Nanoparticles for Selective Cell Membrane Glycan Labeling and Cancer Cell Imaging

Molecular imaging enables in situ visualization of biomolecules in living organisms and creates numerous opportunities for basic biological research and early disease diagnosis. As luminescent probes for molecular imaging, lanthanide-doped upconversion nanoparticles (UCNPs) exhibit superior performance compared to conventional fluorescent dyes in many ways, including high tissue penetration depth and minimized autofluorescence and photobleaching, making them particularly advantageous for imaging analysis. Although various synthesis methods have been reported, the preparation of high quality, water-soluble UCNPs remains challenging. For in situ imaging, glycans on the cell surface are particularly attractive due to their key roles in cellular activity and disease occurrence and development. However, glycan imaging is a challenging task due to their diverse structures and incompatibility with genetically encoded fluorescent tagging techniques. Herein, we report a new type of highly water-soluble, lectin-functionalized core–shell UCNP synthesized by surface-initiated atom transfer radical polymerization (SI-ATRP) for selective cell membrane glycan labeling and cancer cell imaging. SI-ATRP modification results in controlled growth of hydrophilic polymers on the UCNP surface and well-defined core–shell structure, producing UCNPs with improved biocompatibility and intact luminance property. Furthermore, the numerous functional groups on the polymer brush shell provide a large number of binding sites and 3D support for lectin immobilization. The increased loading density and diversified architecture of the immobilized lectins facilitates multivalent binding between the lectins and the glycans on the cell surface and leads to selective labeling of highly metastatic hepatocellular carcinoma cells (HCCHM3) in vitro and successful in vivo imaging of HCCHM3 inoculated mice

MOESM3 of Significantly improved solvent tolerance of Escherichia coli by global transcription machinery engineering

Additional file 3: Figure S3. Recombinant expression of pepB, gapA, sdhB and yfgM in E. coli JM109 and their corresponding knockout strains

MOESM1 of Significantly improved solvent tolerance of Escherichia coli by global transcription machinery engineering

Additional file 1: Figure S1. 2D-PAGE of total proteins of E. coli JM109/pHACM-rpoD WT without solvent