A set of experiments, that employs a luxG-expression plasmid construct (pGhis) and is suitable for an undergraduate laboratory course, is presented. Hexahistidine-tagged protein is expressed in E. coli from pGhis, with the T7 RNA polymerase//ac repressor induction system. Bacteria are lysed ...
Recombinant proteins are extensively utilized as reagents in laboratory experiments and to generate antibody probes for analyzing protein synthesis in cells and in organisms (Peter et al., 2008). Many other functional applications of rDNA are found in industry, food production, human/veterinary ...
Although plasma phospholipid transfer protein (PLTP) has been mainly studied in the context of atherosclerosis, it shares homology with proteins involved in innate immunity. Here, we produced active recombinant human PLTP (rhPLTP) in the milk of new lines of transgenic rabbits. We successfully used...
Transmembrane proteins vary in the number of transmembrane helices (Figure 1), size, overall hydrophobicity, size of domains outside the membrane, and other physical features that might play a role in determining if a transmembrane protein will be expressed at high levels in E. coli. Identifying...
3.2. NGF Protein Biosynthesis After identifying the “Golden Batch” trajectories for the E. coli BL21 biomass growth, multiple fermentation experiments were carried out with the induction of NGF recombinant protein biosynthesis after reaching a specified cell density (Figure 5). Recombinant protein synt...
(TSH). The isolation of a full-length cDNA encoding the commonα subunitdemonstrates that there is a single human gene for this protein, expressed in the placenta for production ofhCGand in the pituitary for the production ofhLH, hFSH, and hTSH. From this, it is concluded that the ...
protein was expressed in anE. colisystem using pET-28a vector (Novagen, USA). Recombinant nanobody fusions against the FliC protein were expressed in anE. colisystem using the pET-25b vector (Novagen, USA). The pMECS vector and M13KO7 helper phage were preserved in our laboratory and were...
eADF4(C16) can be further modified, allowing the protein to be tailored to specific applications26,27. As cells do not attach well to eADF4(C16)28, the aim of this study was to evaluate the interaction of cardiomyocytes with RGD-functionalized eADF4(C16) coatings. For this purpose, we ...
Further experiments were performed, using the endogenous cytoplasmic β-galactosidase as a reporter protein, to determine if PLC caused cell lysis [27]. Our results showed that the percentage of β-galactosidase activity in the culture medium at the time when the extracellular PLC activity had ...
Unit DefinitionOne unit is defined as the amount of enzyme needed to cleave 100 μg of fusion protein in 16 hours to 90 % completion at 5 °C in a buffer containing 50 mM Tris-HCl, pH 7.0, 150 mM NaCl, 1 mM EDTA, and 1 mM DTT. ...