PMC:2682197 / 46439-47173 JSONTXT

{"target":"https://pubannotation.org/docs/sourcedb/PMC/sourceid/2682197","sourcedb":"PMC","sourceid":"2682197","source_url":"http://www.ncbi.nlm.nih.gov/pmc/2682197","text":"Cloning, expression and purification of Rv2623\nThe coding sequence of rv2623 was PCR-amplified from M. tuberculosis Erdman genomic DNA and subcloned into the expression vector pQE80L (Qiagen, Inc.), which encodes an N-terminal His6-tag, producing the plasmid pQE-rv2623 (Text S1). Expression was carried out following isopropyl beta-D-thiogalactoside (IPTG) induction of BL21 E. coli transformed with pQE-rv2623. His6-Rv2623 was then affinity-purified to homogeneity from BL21 cell lysates using Ni-NTA agarose (Qiagen, Inc.) according to the manufacturer's instructions. For crystallization, purified Rv2623 was concentrated to 12 mg/ml using a 10 kDa Molecular Weight Cut Off (MWCO) centrifugal filter (Amicon), and frozen at −80°C.","divisions":[{"label":"Title","span":{"begin":0,"end":46}}],"tracks":[]}