PMC:4236617 / 9417-10264 JSONTXT

{"target":"https://pubannotation.org/docs/sourcedb/PMC/sourceid/4236617","sourcedb":"PMC","sourceid":"4236617","source_url":"http://www.ncbi.nlm.nih.gov/pmc/4236617","text":"Cloning and heterologous expression of the genes encoding L. amylovorus S-layer proteins\nThe expressed slp genes (see above) were amplified by PCR from the chromosomal DNA of the L. amylovorus strains, cloned as NcoI-XhoI –fragments in E. coli DH5αF', sequenced to verify the correct open reading frames, and expressed in E. coli BL21 (DE3) as C-terminal hexahistidine tag-fusions, as described in the pET system manual (Merck KGaA, Darmstadt, Germany) and as previously reported [33]. Recombinant S-layer proteins were purified in the presence of 4 M guanidine hydrochloride (GuHCl) with His Trap HP columns (GE Healthcare, Little Chalfont, UK) according to the manufacturer’s instructions. The pooled protein fractions were dialysed against deionized water overnight at +4°C, centrifuged (15 000 g, 20 min, +4°C) and stored in aliquots at -80°C.","divisions":[{"label":"Title","span":{"begin":0,"end":88}}],"tracks":[{"project":"2_test","denotations":[{"id":"25070625-18828902-12189972","span":{"begin":481,"end":483},"obj":"18828902"}],"attributes":[{"subj":"25070625-18828902-12189972","pred":"source","obj":"2_test"}]},{"project":"MicrobeTaxon","denotations":[{"id":"T330","span":{"begin":58,"end":71},"obj":"1604"},{"id":"T73","span":{"begin":58,"end":71},"obj":"1604"},{"id":"T74","span":{"begin":179,"end":192},"obj":"1604"},{"id":"T75","span":{"begin":322,"end":334},"obj":"511693"},{"id":"T76","span":{"begin":236,"end":250},"obj":"668369"}],"namespaces":[{"prefix":"_base","uri":"http://purl.bioontology.org/ontology/NCBITAXON/"}],"attributes":[{"subj":"T330","pred":"source","obj":"MicrobeTaxon"},{"subj":"T73","pred":"source","obj":"MicrobeTaxon"},{"subj":"T74","pred":"source","obj":"MicrobeTaxon"},{"subj":"T75","pred":"source","obj":"MicrobeTaxon"},{"subj":"T76","pred":"source","obj":"MicrobeTaxon"}]}],"config":{"attribute types":[{"pred":"source","value type":"selection","values":[{"id":"2_test","color":"#93ecb7","default":true},{"id":"MicrobeTaxon","color":"#ec9d93"}]}]}}