Improved mycobacterial protein production using a Mycobacterium smegmatis groEL1ΔC expression strain

<p>Abstract</p> <p>Background</p> <p>The non-pathogenic bacterium <it>Mycobacterium smegmatis </it>is widely used as a near-native expression host for the purification of <it>Mycobacterium tuberculosis </it>proteins. Unfortunately, the Hsp60 chaperone GroEL1, which is relatively highly expressed, is often co-purified with polyhistidine-tagged recombinant proteins as a major contaminant when using this expression system. This is likely due to a histidine-rich C-terminus in GroEL1.</p> <p>Results</p> <p>In order to improve purification efficiency and yield of polyhistidine-tagged mycobacterial target proteins, we created a mutant version of GroEL1 by removing the coding sequence for the histidine-rich C-terminus, termed GroEL1ΔC. GroEL1ΔC, which is a functional protein, is no longer able to bind nickel affinity beads. Using a selection of challenging test proteins, we show that GroEL1ΔC is no longer present in protein samples purified from the <it>groEL1ΔC </it>expression strain and demonstrate the feasibility and advantages of purifying and characterising proteins produced using this strain.</p> <p>Conclusions</p> <p>This novel <it>Mycobacterium smegmatis </it>expression strain allows efficient expression and purification of mycobacterial proteins while concomitantly removing the troublesome contaminant GroEL1 and consequently increasing the speed and efficiency of protein purification.</p

How COVID-19 has changed the unselected medical take: an observational study

INTRODUCTION: COVID-19 has had a profound effect on the NHS. Little information has been published as to how the unselected medical take has been affected. METHODS: We retrospectively reviewed patients who were referred to general medicine during March 2020. We compared clinical outcomes of patients with and without COVID-19. RESULTS: 814 patients were included, comprising 777 unique patients. On average, 26 patients were admitted per day. 38% of admitted patients were suspected of COVID-19, with greater numbers of COVID-19 patients in the second half compared to the first half of the month (p<0.001). Logistic regression analyses showed suspected COVID-19 was an independent predictor for inpatient mortality (odds ratio [OR] = 6.09, p<0.001) and 30-day mortality (OR = 4.66, p<0.001). CONCLUSIONS: COVID-19 patients had worse clinical outcomes and increased healthcare use compared to non-COVID-19 patients. Our study highlights some of the challenges in healthcare provision faced during this pandemic

Proceedings of the 6th International Conference on Modeling and Simulation in Civil Engineering

This conference proceedings contains articles on the various research ideas of the academic community and technical researchers presented at the 6th International Conference on Modeling and Simulation in Civil Engineering (ICMSC 2022). ICMSC 2022 was organized by the Department of Civil Engineering, TKM College of Engineering, Kollam, Kerala, India on December 1-3, 2022. The main aim of this conference is to bring together leading academicians, researchers, technocrats, practitioners, and students to exchange and share their experiences and research outputs on all aspects of Civil Engineering, especially related to the modeling and simulation in Civil Engineering.  Conference Title: 6th International Conference on Modeling and Simulation in Civil EngineeringConference Acronym:  ICMSC 2022Conference Date: 1-3 December 2022Conference Location: IndiaConference Organizer: Department of Civil Engineering, TKM College of Engineering, Kollam, Kerala, Indi