Abstract: The present application relates to a modified Prp43 helicase and use thereof. The Prp43 helicase has enhanced ATP hydrolytic or unwinding activity due to introduction of mutations and/or introduction of an auxiliary protein, and can remain binding to the target polynucleotide for a long period of time, thereby allowing the enzyme to control the rate of movement of the polynucleotide continuously and stably at a suitable rate required for sequencing. Thus, the modified or engineered Prp43 helicase mutant of the present application allows for the control of movement of a target polynucleotide in a more advantageous manner, which can be used for nanopore sequencing.

Abstract: The present application provides a Pif1-like helicase and the use thereof, specifically a modified Pif1-like helicase, a construct comprising the Pif1-like helicase, and its use in characterising a target polynucleotide or controlling the movement of a target polynucleotide through a pore. The present application also provides a method for characterising the target polynucleotide or controlling the movement of the target polynucleotide through the pore. The Pif1-like helicase of the present application can effectively control the movement of the polynucleotide through the pore.

Abstract: Provided are a Phi29 DNA polymerase mutant with improved thermal stability and an use thereof in sequencing. The phi29 DNA polymerase mutant is represented by A) or B) below: the DNA polymerase mutant represented by the A) is a protein having DNA polymerase activity that is obtained by modifying at least one amino acid residue(s) in the following six sites in a phi29 DNA polymerase amino acid sequence: the 97-th, 123-th, 217-th, 224-th, 515-th, and 474-th sites; the DNA polymerase mutant represented by the B) is a protein having DNA polymerase activity that is derived from the A) by adding a tag sequence to a terminal of the amino acid sequence of the protein represented by the A).

Abstract: The present disclosure relates to a reverse transcriptase and an application thereof. The reverse transcriptase has mutation sites such as R450H compared with the wild-type M-MLV reverse transcriptase. The reverse transcriptase has increased polymerase activity, improved thermal stability, and reduced RNase H activity.

Abstract: Provided are a group of phi29 DNA polymerase mutants having increased thermal stability and use thereof. The phi29 DNA polymerase mutants are proteins obtained by performing point mutation A and/or point mutation B and/or point mutation C on phi29 DNA polymerase, the point mutation A meaning that an amino acid residue M at position 97 of the phi29 DNA polymerase is mutated to other amino acid residue, the point mutation B meaning that an amino acid residue L at position 123 of the phi29 DNA polymerase is mutated into other amino acid residue, and the point mutation C meaning that an amino acid residue E at position 515 of the phi29 DNA polymerase is mutated to other amino acid residue. The stability of the phi29 DNA polymerase mutants is higher than that of a wild-type phi29 DNA polymerase.

Abstract: Provided are a phi29 DNA polymerase mutant with increased thermo stability, a method for preparing the mutant, the use of the mutant, and a method for increasing the stability of the phi29 DNA polymerase.

Abstract: Provided are a phi29 DNA polymerase and an encoding gene and an application thereof. The phi29 DNA polymerase is C1) or C2): C1) is a protein with DNA polymerase activity obtained by substituting at least one of the 58th, 61st, 94th, 96th, 119th, and 155th amino acid residues in the amino acid sequence of a wild type phi29 DNA polymerase as shown in SEQ ID NO: 2 in the sequence listing; and C2) is a fusion protein obtained by linking a label to the N-terminus and/or C-terminus of the protein represented by C1). A 3?-5?exonuclease of the phi29 DNA polymerase has activity lower than that of the wild type phi29 DNA polymerase, and can efficiently and continuously synthesize DNA during amplification and sequencing.

Abstract: Provided are a group of phi29 DNA polymerase mutants having increased thermal stability and use thereof. The phi29 DNA polymerase mutants are proteins obtained by performing point mutation A and/or point mutation B and/or point mutation C on phi29 DNA polymerase, the point mutation A meaning that an amino acid residue M at position 97 of the phi29 DNA polymerase is mutated to other amino acid residue, the point mutation B meaning that an amino acid residue L at position 123 of the phi29 DNA polymerase is mutated into other amino acid residue, and the point mutation C meaning that an amino acid residue E at position 515 of the phi29 DNA polymerase is mutated to other amino acid residue. The stability of the phi29 DNA polymerase mutants is higher than that of a wild-type phi29 DNA polymerase.

Abstract: Provided are a phi29 DNA polymerase mutant with increased thermo stability, a method for preparing the mutant, the use of the mutant, and a method for increasing the stability of the phi29 DNA polymerase.

Abstract: Provided are a phi29 DNA polymerase and an encoding gene and an application thereof. The phi29 DNA polymerase is C1) or C2): C1) is a protein with DNA polymerase activity obtained by substituting at least one of the 58th, 61st, 94th, 96th, 119th, and 155th amino acid residues in the amino acid sequence of a wild type phi29 DNA polymerase as shown in SEQ ID NO: 2 in the sequence listing; and C2) is a fusion protein obtained by linking a label to the N-terminus and/or C-terminus of the protein represented by C1). A 3?-5?exonuclease of the phi29 DNA polymerase has activity lower than that of the wild type phi29 DNA polymerase, and can efficiently and continuously synthesize DNA during amplification and sequencing.