A novel pro-apoptosis protein PNAS-4 from xenopus laevis: cloning, expression, purification, and polyclonal antibody production [ J ]. Biochemistry ( Moscow), 2007,72:664-671.Fei Yan,Meilin Qian,Fan Yang,Feng Ca
The inclusion bodies were isolated by high-speed centrifugation, washed, and resuspended in the denaturation buffer. The dissolved pellet was subjected to incubation for 2 h, followed by high-speed centrifugation. The supernatant was loaded onto a pre-equilibrated Ni-NTA column, and the bound ...
SDS-PAGE analysis (Supplementary information, Figure S2) showed that all variants were expressed in soluble form, we did not observed the formation of inclusion bodies for wild-type OxdB or any of its variants. The relative expression level compared to the wild-type enzyme was in overall ...
However, the ClyA-SS mutants from this study have challenges in terms of low water solubility and expression into inclusion bodies. As a result, directed evolution was further used to overcome these challenges to design two new mutants ClyA-CS (L99Q, E103Q, F166Y, C285S, K294R) and ...
10.1073/pnas.93.3.1314. Article PubMed PubMed Central Google Scholar Querfurth HW, Suhara T, Rosen KM, McPhie DL, Fujio Y, Tejada G, Neve RL, Adelman LS, Walsh K: Beta-amyloid peptide expression is sufficient for myotube death: implications for human inclusion body myopathy. Mol Cell ...
7). We proposed that the poor solubilities of these three mutants may due to their decreased folding stabilities, therefore leading to high tendency to precipitate as inclusion bodies (data not shown). As will be shown later by GdmHCl-induced denaturation experiments, these three mutations indeed...
Human PrP23-231 (rhPrP) was expressed inE. coliBL21(DE3) cells and purified from the inclusion bodies on a Ni–NTA column as described [28]. Briefly, protein expression was induced by culturing bacteria in M9 minimal medium containing 1 mM isopropil-β-D-1-tiogalattopiranoside (IPTG, ...
coli is largely insoluble and must be purified from inclusion bodies [27]. This procedure requires denaturation and refolding steps, involves harsh chemical treatments, and often yields low amounts of correctly folded protein. A subsequent report demonstrated that only the laforin CBM was soluble ...
The rate of fluorescence acquisition of YFP renatured from inclusion bodies would inform us of the kinetics of de novo maturation, including the step of cyclization of an internal tripeptide (Reid, B. G. et al (1997) Biochemistry 36, 6786-6791). However, SEYFP and SEYFP-F46L were hardly...
When a portion of the culture broth was sampled and observed under a phase-contrast microscope (magnification, 1000 to 1500), inclusion bodies in the form of granules were found in thinly elongated E. coli cells. Thereafter, the culture broth was subjected to centrifugation (8000 rpm, 5 minute...