Cloning, expression and characterization of the Sau3AI restriction and modification genes in Staphylococcus carnosus TM300

S Seeber, C Kessler, F Götz - Gene, 1990 - Elsevier
S Seeber, C Kessler, F Götz
Gene, 1990Elsevier
The genes encoding the restriction enzyme (ENase) and modification enzyme (MTase) of
Staphylococcus aureus 3A (recognition sequence 5′-GATC-3′) have been cloned in
Staphylococcus carnosus TM300 using the vector pCA44. Clones carrying both genes were
isolated from DNA libraries prepared with Mbo I+ Bam HI. The DNA region encoding Mˇ
Sau3AI was subcloned on a 3.66-kb EcoRV fragment in vector pT181mcs. Plasmids purified
from the clones were resistant to digestion with Sau3AI, indicating that the sau3AIM gene …
The genes encoding the restriction enzyme (ENase) and modification enzyme (MTase) of Staphylococcus aureus 3A (recognition sequence 5′-GATC-3′) have been cloned in Staphylococcus carnosus TM300 using the vector pCA44. Clones carrying both genes were isolated from DNA libraries prepared with Mbo I+ Bam HI. The DNA region encoding Mˇ Sau3AI was subcloned on a 3.66-kb EcoRV fragment in vector pT181mcs. Plasmids purified from the clones were resistant to digestion with Sau3AI, indicating that the sau3AIM gene was expressed and the product was functional in S. carnosus. Cell lysates of clones with both activities encoded on plasmid pSEM7, cut DNA with the same pattern as Sau3AI, showing that the sau3AIR gene was also expressed and the ENase was functional in S. carnosus. Sequence analysis shows that both genes are transcribed in the same direction and encode polypeptides with calculated M r s of 56477 for Rˇ Sau3AI and 47 300 for Mˇ Sau3AI. Efforts to clone one or both genes in Escherichia coli have so far failed.
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