fragments of DNA from NCIB 8565, a high producer of an endo-1,3-1,4-β-D-glucanase, were ‘shot-gun’ cloned in the plasmid vector pBR325. A 3.5 kb insert, carrying single restriction sites for I, II, I, I and II, was shown to direct the synthesis of β-glucanase in K12. Enzyme activity was demonstrated in extracellular fractions of harbouring the β-glucanase gene; however, the largest proportion (> 50%) of total enzyme activity was periplasmic in location. β-Glucanase activity and cellular location were independent of the orientation of the 3.5 kb fragment in pBR325.


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