Method for cloning and expression of BsaI restriction endonuclease and BsaI methylase in E. coli

• US 20030186363A1
• Filed: 03/26/2002
• Published: 10/02/2003
• Est. Priority Date: 03/26/2002
• Status: Active Grant

• Current Assignee: New England Biolabs Incorporated
• Original Assignee: New England Biolabs Incorporated
• Inventors: Zhu, Zhenyu, Xu, Shuang-Yong
• Granted Patent: US 6,723,546 B2
• Time in Patent Office: Days
• US Class Current: 435/69.1
• CPC Class Codes: C12N 9/1007 Methyltransferases (general) (2.1.1.)
• CPC Class Codes: C12N 9/22 Ribonucleases RNAses, DNAses catalytic nucleic acids C12N15/113

Abstract

The present invention relates to recombinant DNA which encodes the BsaI restriction endonuclease as well as BsaI methylase, expression of BsaI restriction endonuclease and BsaI methylase in E. coli cells containing the recombinant DNA, and purification of BsaI restriction endonuclease to near homogeneity.

Claims

1. Isolated DNA coding for the BsaI restriction endonuclease, wherein the isolated DNA is obtainable from Bacillus stearothermophilus 6-55.

2. A recombinant DNA vector comprising a vector into which a DNA segment encoding the BsaI restriction endonuclease gene has been inserted.

5. A host cell transformed by the vector of claims 2 or 4.

6. A method of producing recombinant BsaI restriction endonuclease comprising culturing a host cell transformed with the vector of claims 2 or 4 under conditions suitable for expression of said endonuclease and methylase.

3. Isolated DNA encoding the BsaI restriction endonuclease and BsaI methylase, wherein the isolated DNA is obtainable from ATCC No. ______.

4. Vectors that comprise the isolated DNA of claim 3.