Please use this identifier to cite or link to this item: https://doi.org/10.3390/pr9040629
Title: High Throughput Expression Screening of Arabinofuranosyltransferases from Mycobacteria
Authors: Rodrigues, Jose
Almeida, Vanessa T
Rosario, Ana L
Tan, Yong Zi 
Kloss, Brian
Mancia, Filippo
Archer, Margarida
Keywords: Science & Technology
Technology
Engineering, Chemical
Engineering
membrane proteins
overexpression in E. coli
protein purification
high-throughput protocol
Arabinofuranosyltransferases
Mycobacteria
MEMBRANE-PROTEIN OVEREXPRESSION
ESCHERICHIA-COLI
BIOSYNTHESIS
ARABINOGALACTAN
PURIFICATION
STRATEGY
Issue Date: 1-Apr-2021
Publisher: MDPI
Citation: Rodrigues, Jose, Almeida, Vanessa T, Rosario, Ana L, Tan, Yong Zi, Kloss, Brian, Mancia, Filippo, Archer, Margarida (2021-04-01). High Throughput Expression Screening of Arabinofuranosyltransferases from Mycobacteria. PROCESSES 9 (4). ScholarBank@NUS Repository. https://doi.org/10.3390/pr9040629
Abstract: Studies on membrane proteins can help to develop new drug targets and treatments for a variety of diseases. However, membrane proteins continue to be among the most challenging targets in structural biology. This uphill endeavor can be even harder for membrane proteins from Mycobacterium species, which are notoriously difficult to express in heterologous systems. Arabino-furanosyltransferases are involved in mycobacterial cell wall synthesis and thus potential targets for antituberculosis drugs. A set of 96 mycobacterial genes coding for Arabinofuranosyltransferases was selected, of which 17 were successfully expressed in E. coli and purified by metal-affinity chro-matography. We herein present an efficient high-throughput strategy to screen in microplates a large number of targets from Mycobacteria and select the best conditions for large-scale protein production to pursue functional and structural studies. This methodology can be applied to other targets, is cost and time effective and can be implemented in common laboratories.
Source Title: PROCESSES
URI: https://scholarbank.nus.edu.sg/handle/10635/227198
ISSN: 22279717
DOI: 10.3390/pr9040629
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