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Molecular Biology

Method-for-producing-the-ACCI-restriction-endonuclease-and-methylase

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Method for producing the ACCI restriction endonuclease and methylase

Details

Inventors: Chen, Shu-zi; Wilson, Geoffrey G.;
Assignee: New England Biolabs, Inc. (Beverly, MA)
Primary Examiner: Weimar; Elizabeth C.
Assistant Examiner: Patterson, Jr.; Charles L.
Attorney, Agent or Firm: Williams; Gregory D., Conlin; David G.

The present invention is directd to a method for cloning and producing the AccI restriction endonuclease by (1) introducing the restriction endonuclease gene from into a host whereby the restriction gene is expressed; (2) fermenting the host which contains the plasmid encoding and expressing the AccI restriction endonuclease activity, and (3) purifying the AccI restriction endonuclease from the fermented host which contains the plasmid encoding and expressing the AccI restriction endonuclease activity.

DETAILED DESCRIPTION In accordance with the present invention there is provided a clone containing the genes for the AccI restriction endonuclease and modification methylase derived from Acinetobacter calcoaceticus, as well as related methods for the production of the enzymes.
More specifically, this invention relates to clones which express the restriction endonuclease AccI, which recognizes the DNA sequence 5' .
.
.
GT(AC)(GT)AC .
.
.
3' and cleaves after the first T.
The preferred method for cloning this enzyme comprises forming a library containing the DNA from Acinetobacter calcoaceticus, isolating those clones which contain DNA coding for the AccI modification methylase and screening these to identify those that also contain the AccI restriction endonuclease gene.

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