Bst Preliminary Patent Landscape

Hi All

Please find below preliminary information of the Bst Patent Landscape based on US patents.
This is a starting point - please help to improve it if you have knowledge or time to research!

Jenny


Bst Patent Landscape

Caveat: this is based on USPTO and EPO data only, the patents mentioned as live may not have been filed in your jurisdiction. Also it does not constitute legal advice and may well be wrong in parts. Please help to improve it!

Bst is a DNA Pol I strand-displacing polymerase from Bacillus stearothermophilus. The full length (FL) Bst DNA polymerase is 876 amino acid residues and has 5β€²-3β€² endonuclease activity but not 3β€²-5β€² exonuclease activity. The large fragment (LF) of Bst DNA polymerase lacks both 5β€²-3β€² exonuclease activity and 3β€²-5β€² exonuclease activity and is only 587 amino acid residues with 289 amino acids being deleted from the N-terminal end. The large fragment Bst DNA polymerase have been found to be useful for isothermal amplification techniques and DNA sequencing.

Patent for Production of Bst Full Length [Expired]

US 6066483 A (Priority Apr 1, 1994)
Composition and methods for the expression of recombinant DNA polymerase enzymes derived from Bacillus stearothermophilus. The present invention also concerns methods for purifying recombinant Bst DNA polymerase enzymes, compositions containing the purified enzymes in a form suitable for conducting biochemical reactions, and methods for using the purified enzymes.

Patent for production of Bst LF [Expired]

US 5830714 A (Priority Apr 17, 1996)
Biologically Active Fragment Of Bacillus Stearothermophilus Dna Polymerase
The present invention is directed to an isolated and purified DNA encoding a biologically active fragment of a thermostable, full length DNA polymerase I enzyme of Bacillus stearothermophilus. More particularly, the invention is directed to a DNA encoding an approximately 66,000 dalton DNA polymerase that lacks 273 amino acids from the N-terminus of the approximately 96,000 dalton B. stearothermophilus DNA polymerase I, and to the protein encoded thereby which has been designated the B. stearothermophilus DNA polymerase I exo- fragment.

US 5814506 A (Earliest Priority: Aug 02 1995)
Over-expression And Purification Of A Truncated Thermostable Dna Polymerase By Protein Fusion
The present invention relates to the cloning and purification of a thermostable DNA polymerase, Bst polymerase I from Bacillus stearothermophilus. More specifically, it provides a novel method for producing a truncated Bst polymerase using recombinant DNA and protein fusion techniques.

Production of Bst with Reduced 3’- To -5’ Exonuclease Activity [Expired]

US 6013451 A 9 (Earliest Priority: Apr 10 1997,)
Bacillus Stearothermophilus Dna Polymerase I (klenow) Clones Including Those With Reduced 3’- To -5’ Exonuclease Activity
Disclosed and claimed are isolated nucleic acid molecules encoding Bacillus stearothermophilus DNA polymerase (DNApolI), including the structural gene for DNApolI, such as DNApolI genes having insertions, deletions, inactivation, or mutations at the 5’ end thereof and thus encode Bst polymerase I enzymes which lack or have reduced 3’-5’ exonuclease activity, as well as methods for making and using such nucleic acid molecules and such polymerases.

Use of Bst for reverse transcription [Expired]

WO 2000/071739 A1 (Earliest Priority: May 22 1999)
No USPTO grant
The present invention is directed to a thermostable DNA polymerase from Bacillus stearothermophilus for use in reverse transcription and/or reverse transcriptase-polymerase chain reaction (RT-PCR), where said DNA polymerase shows magnesium ion dependent reverse transcriptase activity and in the substantial absence of manganese ions.

US 7094539 B2(Earliest Priority: Mar 02 2000)

The present invention relates to reverse transcription of RNA, and in particular to reverse transcription by thermostable DNA polymerases. Thermoactinomyces vulgaris and Bacillus stearothermophilus possess reverse transcriptase activity in the presence of magnesium or manganese ions. Methods, compositions, and kits for reverse transcription and RT-PCR are also provided.

US 6949634 B2 (Earliest Priority: Nov 27 1998)

A method for synthesizing cDNA characterized by performing a reverse transcription reaction in the presence of an enzyme having a reverse transcriptional activity and another enzyme different from the former one which as a 3’-5’ exonuclease activity.

Improvements to Bst [Expired]

US 9840698 B2 (Earliest Priority: Jun 18 2010)
Dna Polymerases With Increased 3β€²-mismatch Discrimination
Disclosed are mutant DNA polymerases having increased 3β€²-mismatch discrimination relative to a corresponding, unmodified polymerase. The mutant polymerases are useful in a variety of disclosed primer extension methods. Also disclosed are related compositions, including recombinant nucleic acids, vectors, and host cells, which are useful, e.g., for production of the mutant DNA polymerases.

US 6818431 B1 (Earliest Priority: Oct 18 1995)
Dna Polymerase Having Ability To Reduce Innate Selective Discrimination Against Fluorescent Dye-labeled Dideoxynucleotides
The invention relates to genetic modification of DNA polymerase to reduce innate selective sequence-related discrimination against incorporation of fluorescent dye-labeled ddCTP and ddATP in the enzymatic reaction for preparation of samples for automated fluorescent dye-labeled terminator DNA sequencing. The modified DNA polymerases are more resistant to heat inactivation and are more effective in dideoxynucleotide incorporation than current DNA polymerases.

US 5834253 A (Earliest Priority: May 03 1996)
Bacillus Stearothermophilus Dna Polymerase With Proof-reading 3’-5’ Exonuclease Activity
The invention relates to DNA polymerases which are capable of proofreading 3’-5’ exonuclease activity during DNA sequencing of a DNA strand, such that the DNA polymerase functions to excise mismatched nucleotides from the 3’ terminus of the DNA strand at a faster rate than the rate at which the DNA polymerase functions to remove nucleotides matched correctly with the nucleotides of the template, and which DNA polymerase does not exhibit 5’-3’ exonuclease activity. The invention also relates to isolated and cloned DNA sequences derived from the Bacillus stearothermophilus thermostable DNA polymerase, as well as the expressed polymerase itself.

Improvements to Bst [Active US Patent - check local]

US 9109226 B2 (Earliest Priority: Sep 01 2011)
Aptamer inhibition - Synthetic Nucleic Acids For Polymerization Reactions
Compositions and methods are provided for inhibiting a polymerase from replicating non target DNA at a temperature below the amplification reaction temperature. The inhibitor is a synthetic nucleic acid which is single stranded but folds to form at least one double stranded region designed to melt at a temperature which is lower than the amplification reaction temperature, and at least one single stranded region where the single stranded region at the 5β€² end contains at least one uracil or inosine and optionally a sequence at the 3β€² end contains one or more derivative nucleotide or linkages.

US 9738877 B2 (Earliest Priority: Jul 28 2011)
Dna Polymerases With Improved Activity
Disclosed are DNA polymerases having increased reverse transcriptase efficiency relative to a corresponding, unmodified polymerase. The polymerases are useful in a variety of disclosed primer extension methods. Also disclosed are related compositions, including recombinant nucleic acids, vectors, and host cells, which are useful, e.g., for production of the DNA polymerases.

Bst 2.0 from NEB [Active US Patent - check local]

US 9157073 B1 (Earliest Priority: Aug 31 2012)
Dna Polymerases
Novel proteins having DNA polymerase are described which have utility in amplification reactions and have improved properties over Bst polymerase such as for example enhanced reverse transcriptase activity.

EP 2751264 B1 (Earliest Priority: Sep 01 2011)
Compositions And Methods Relating To Variant Dna Polymerases And Synthetic Dna Polymerases
Compositions of novel polymerase variants and methods of identifying, making and using these novel polymerases are described. The variants have been shown to have advantageous properties such as increased thermostability, deoxyuridine nucleoside triphosphate tolerance, salt tolerance, reaction speed and/or increased reverse transcriptase properties. Uses for these improved enzymes have been demonstrated in isothermal amplification such as LAMP. Enhanced performance resulting from the use of these variants in amplification has been demonstrated both in reaction vessels and in dedicated automated amplification platforms.

Specific Improvements to Bst LF

M316R = increased salt tolerance in the range of 20 mM - 200 mM monovalent salt (EP 2751264 B1 - active patent)

G310L = increased efficiency. Ma, Yi, et al. β€œEnhancement of polymerase activity of the large fragment in DNA polymerase I from geobacillus stearothermophilus by Site-Directed Mutagenesis at the active site.” BioMed research international 2016 (2016) [patent status unknown]

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