{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,3]],"date-time":"2026-10-03T08:39:42Z","timestamp":1791016782918,"version":"4.1.0"},"reference-count":27,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2010,12,23]],"date-time":"2010-12-23T00:00:00Z","timestamp":1293062400000},"content-version":"vor","delay-in-days":843,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biotech and App Biochem"],"published-print":{"date-parts":[[2008,9]]},"abstract":"<jats:p>We have previously shown the cyanobacterium <jats:italic>Anabaena<\/jats:italic> sp. PCC 7120 to be a suitable host for the production of isotopically labelled recombinant proteins using the nitrate\u2010inducible <jats:italic>nir<\/jats:italic> expression system. However, the expression of toxic proteins such as oncoproteins proved to be difficult, as expression levels decreased shortly after induction, while growth continued. To overcome this limitation, we have developed a method of auto\u2010induction of the <jats:italic>nir<\/jats:italic> promoter in which cells are grown to high cell density in a bioreactor in the presence of ammonium and nitrate. Since ammonium is the preferred nitrogen source and acts as a repressor of the <jats:italic>nir<\/jats:italic> promoter, induction occurs only when the ammonium had been depleted. Using this novel auto\u2010induction method, both oncoproteins E6 and gankyrin were expressed at high levels in a folded conformation and were shown to be biologically active after purification. Furthermore, under similar conditions of growth in auto\u2010inducing medium, the use of <jats:sup>15<\/jats:sup>N\u2010 and <jats:sup>13<\/jats:sup>C\u2010labelled mineral salts yielded isotopic enrichment of these proteins at levels above 95%, making them suitable for NMR\u2010based structural analysis in a cost\u2010effective manner.<\/jats:p>","DOI":"10.1042\/ba20070276","type":"journal-article","created":{"date-parts":[[2008,5,12]],"date-time":"2008-05-12T13:18:15Z","timestamp":1210598295000},"page":"53-61","source":"Crossref","is-referenced-by-count":6,"title":["Automated overexpression and isotopic labelling of biologically active oncoproteins in the cyanobacterium <i>Anabaena<\/i> sp. 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