Table 1.

Biological activity of SPase II genes from various bacteria in E. colia

PlasmidGeneGrowth at the following Glm concn (μg ml−1)Complementation growth of E. coli Y815b
0102550200
None+++
pUC19+++NT
pQE9+++
SUPERCOS-1+++NT
pPY06c taG +++NT
pPYCA111c taG +++NT
pPYMXSP-IId 6 His-taG +++++±
pPYECSP-IIe 6 His-lsp ++++++
pKSNTNTNTNTNT
pKSA11f B. subtilis lsp NTNTNTNTNT+
  • a Bacterial cultures (E. coliTG1 and its transformants) were grown in LB containing 0 to 200 μg of Glm ml−1 and 100 μg of ampicillin ml−1 (if required) at 30, 37, and 42°C. The results (+, growth; ±, limited growth; and −, no growth) were the same for all temperatures tested, and therefore, the temperature is not indicated. The same experiment was also performed with E. coli Y815 and its transformants, except that Glm resistance was determined only at 30°C. NT, not tested.

  • b Complementation growth on LB or AB3 plates.

  • c taG was cloned with its own regulatory elements.

  • d taG was cloned under pQE9 regulatory elements.

  • e E. coli lsp was cloned under pQE9 regulatory elements.

  • f B. subtilis ileS-lsp-pyrR operon was cloned in pBluescript KS(+) as described elsewhere (28).