Cloning and expression in XXXX of the gene for an Arthrobacter beta-(1----3)-glucanase.

@entity375 @entity3208 @entity2975 @entity526
When inserted in the correct orientation at the BamHI site of plasmid YRp7, an 8.6-kilobase BamHI fragment of Arthrobacter sp. strain YCWD3 DNA gave @entity526 HB101 cells harboring the recombinant plasmid pBX20 the ability to lyse @entity375 cell walls or @entity375 glucan in agar medium. An extract of the transformed @entity526 cells contained an endo-beta-(1----3)-glucanase with the same activity pattern as that of glucanase I produced by Arthrobacter sp. strain YCWD3. Although part of the glucanase activity was contributed by apparently defective molecules, two protein species were found which had high lytic activity on @entity375 cell walls and adsorbed to microcrystalline cellulose, and both had a single constituent polypeptide with a molecular weight of about 55,000, as determined by @entity3208 - @entity2975 gel electrophoresis. In these properties the protein species were indistinguishable from those glucanase I protein species of Arthrobacter sp. strain YCWD3 which we believe are nearly the intact molecule. We conclude that the cloned fragment of Arthrobacter sp. strain YCWD3 DNA contains the structural gene for glucanase I. A recombinant plasmid obtained by subcloning a PstI fragment of pBX20 into pBR322 caused the transformed @entity526 cells to produce apparently defective glucanase molecules only. This observation serves as additional supporting evidence for our conclusion.
