Loading patent…
AssigneeNew England Biolabs, Inc.
InventorsRichard J. Roberts, Devon R. Byrd, Richard D. Morgan, Jay Patti, Christopher J. Noren
Estimating the term…
A method is provided for identifying a restriction endonuclease, which includes the steps of (a) screening a target DNA sequence for the presence of known methylase sequence motifs, (b) identifying any open reading frames which lie close to the methylase sequence motifs screened in step (a), and (c) assaying the protein products of these open reading frames for restriction endonuclease activity. Methods for identifying isoschizomers of known restriction endonucleases, which isoschizomers possess a desired physical property, such as thermostability, are also provided by the present invention, as are several novel restriction endonucleases isolated from M. jannaschii, MjaIII and MjaIV. Additionally, a gene was identified that encoded a previously observed endonuclease activity, designated MjaII. Also provided by the present invention are vectors suitable for cloning a DNA sequence encoding a cytotoxic protein, via independent transcription promotors which may be selectively controlled by several conditions. A method for producing these cytotoxic proteins using such vectors is also provided, as are stable clones of PacI and NlaIII.
A method for identifying a restriction endonuclease comprising the steps of:
A substantially pure restriction endonuclease MjaIV obtainable from M. jannaschii, said endonuclease recognizing the following base sequence in double-stranded deoxyribonucleic acid molecules:
Isolated DNA coding for the MjaIV restriction endonuclease, wherein the isolated DNA is obtainable from Methanococcus jannaschii.
A recombinant vector comprising a vector into which DNA coding for MjaIV restriction endonuclease has been inserted.
Isolated DNA coding for the MjaII restriction endonuclease, wherein the isolated DNA is obtainable from M. jannaschii.
A recombinant vector comprising a vector into which DNA coding for MjaII restriction endonuclease has been inserted.
A substantially pure MjaIII restriction endonuclease obtainable from M. jannaschii, which recognizes the following base sequence in double-stranded deoxyribonucleic acid molecules:
Isolated DNA coding for the MjaIII restriction endonuclease, wherein the isolated DNA is obtainable from M. jannaschii.
A recombinant vector comprising a vector into which DNA coding for MjaIII restriction endonuclease has been inserted.
A method of identifying an isoschizomer of a known restriction endonuclease, said isoschizomer possessing a desired physical property, said method comprising the steps of:
A vector suitable for cloning a DNA sequence encoding a cytotoxic protein wherein, said vector comprises at least a first and a second transcription promotor and is adapted to accept said DNA sequence insert and wherein said first and said second transcription promotors are independently controllable.
The vector of claim 33, wherein said first transcription promotor enables anti-sense strand transcription and said second transcription promotor enables sense strand transcription.
A stable recombinant vector encoding R.NlaIII, said vector comprising pLT7-nlaIIIR, wherein said vector does not encode M.NlaIII.
Isolated DNA coding for the PacI restriction endonuclease, wherein the isolated DNA is obtainable from ATCC Accession No. 55044.
A recombinant DNA vector comprising a vector into which a DNA segment coding for PacI endonuclease produced by Pseudomonas alcaligenes has been inserted.
A substantially pure MjaV restriction endonuclease obtainable from M. jannaschii, which recognizes the following base sequence in double-stranded deoxyribonucleic acid molecules:
Isolated DNA coding for the MjaV restriction endonuclease, wherein the isolated DNA is obtainable from M. jannaschii.
A recombinant vector comprising a vector into which DNA coding for MjaV restriction endonuclease has been inserted.
Dependencies are inferred from the claim text.