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Cloning - DOAJ

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Last Updated: 19 June 2022

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Cloning, expression and purification of fructosyl amino acid oxidase (FAOX) in Escherichia Coli

"Introduction: The serum HbA1c level is an indicator of the average blood sugar level in the last three months. Using assays involving the enzyme fructosyl amino acid oxidase oxidase, HbA1c can be determined. DH5 (u03b1 and BL21 were used as cloning and expression hosts, respectively, according to the E. coli strains. SDS-PAGE analyzed the protein expression profile. Glutathione Sepharose 4 Fast Flow sonicated and purified the cell pellet, which was sonicated and purified. Using high-performance anion exchange chromatography with pulsed amperometry analysis, the catalytic activity of GST/FAOX-TE with fructosyl valine was established. Conclusions: The fusion protein was successfully expressed in Escherichia coli using the plasmid pGEX-4T3 and purified to 93%. Recombinant GST/FAOX-TE was reported to be operational on fructosyl vain. Conclusions: Active GST/FAOX-TE was successfully expressed in E. coli BL21 and purified, and it was subsequently used for future biosensors for fructosyl valine quantification.

Source link: https://doi.org/10.46223/HCMCOUJS.tech.en.11.1.1238.2021


Molecular cloning and characterization of the porcine prostaglandin transporter ( SLCO2A1 ): evaluation of its role in F4 mediated neonatal diarrhoea

Based on a positional candidate gene approach study, we propose the molecular cloning and characterization of the porcine ortholog in order to determine its potential role in F4 enterotoxigenic E. coli-mediated neonatal diarrhoea. " In 15% of all non-coding sequences, repeat sequences were discovered in 15%. In 5 F4ab/ac receptor positive and 5 F4ab/ac receptor negative pigs, the promoter region and the exons of SLCO2A1 were resequenced. Conclusion The molecular cloning and characterization of porcine SLCO2A1 not only contributes to the already existing body of the transporter, but also allows research on porcine prostaglandin related processes/deficiencies as patient and/or model. SLCO2A1 will most likely be excluded as a receptor for F4 bacteria because no phenotype-related variations could be discovered in the gene sequence nor in its jejunal transcription profile of F4ab/negative pigs.

Source link: https://doi.org/10.1186/1471-2156-10-64


Molecular cloning, expression, and characterization of four novel thermo-alkaliphilic enzymes retrieved from a metagenomic library

"Abstract Background: The Enzyme discovery is a promising step toward assisting in the deconstruction of recalcitrant plant biomass in an industrial process. " In Escherichia coli, eight glycosyl hydrolase family candidate genes were selected from metagenomes of wheat straw-degrading microbial confederation microbial connegation using molecular cloning and subsequent gene expression studies. The corresponding source fosmids, u03b2-galactosidase- and u03b2-xylosidase-like proteins 1 and 2 were identified as being responsible for previously discovered thermo-alkaliphilic glycosidase enzyme activity in extracts of E. coli containing the respective source fosmids. U00b0C and pH range 8. 0-10. 0 were observed with both pNP-Xyl and pNP-Ara in the temperature range 40 to u201310. 0 in the temperature range 40–u201310. 0. The presence of a potential u201cGGDEF_u201d catalytic site, encoding u03b1-glucosidase activity, was revealed by a u201cgDEFu201d catalytic site, encoding a u201d'sidase activity, revealed the presence of a u201cgDEFu201d catalytic site, encoding u203b1-u201d encoding a u201cGMDEFu201d catau201d site encoding u201cu201cu201d u201cu201c au201cg201d u201cg201cu201cggDEFu201du201du201d encoding u201cu201cu201cu201du201cGMDEFu201cu201du201cu202cu201cGMDEFu201c.

Source link: https://doi.org/10.1186/s13068-017-0808-y

* Please keep in mind that all text is summarized by machine, we do not bear any responsibility, and you should always check original source before taking any actions

* Please keep in mind that all text is summarized by machine, we do not bear any responsibility, and you should always check original source before taking any actions