Mgi Tech Co., Ltd.

Bgi Shenzhen

Yuliang Dong

Wenwei Zhang

Chongjun Xu

Yue Zheng

Snezana Drmanac

Zhougang Zhang

Lin Wang

Yujun Zhou

Fen Liu

Huanhuan Liu

Xing Liu

Xi Zhang

Lili Zhai

Provided is a recombinant KOD polymerase, which is the following A) or B): the polymerase shown in A) is a protein having DNA polymerase activity that is obtained by modifying amino acid residues in at least one of the following 18 positions in a wild-type KOD DNA polymerase amino acid sequence: 675th, 385th, 710th, 674th, 735th, 736th, 606th, 709th, 347th, 349th, 590th, 676th, 389th, 589th, 680th, 384th, 496th and 383rd; the polymerase described by B) is a protein having DNA polymerase activity that is derived from A) by adding a tag sequence to an end of the amino acid sequence of the protein shown in A).

Recombinant KOD polymerase

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.

Phi29 DNA polymerase mutant having increased thermal stability and use thereof

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.

PHi29 DNA polymerase mutant with increased thermostability

Disclosed in the present disclosure is a recombinant DNA polymerase. The recombinant DNA polymerase is any one selected from: A) a protein, having amino acid modifications at positions 408, 409 and 485, and at least one of amino acid modification(s) at positions 53, 59, 199, 243, 526, 558, 613, 641, 671, 673, 674, 692 and 709 compared to the amino acid sequence of a wild-type KOD DNA polymerase; B) a protein derived from the protein in A), formed by deleting amino acids 1 to 29 from a C-terminus of the protein in A) and keeping the remaining amino acids unchanged; and C) a protein derived from the protein in A) or B), formed by connecting a tag to the N-terminus or C-terminus of the amino acid sequence of the protein in A) or B), wherein the protein in A), B) and C) has DNA polymerase activity.

DNA polymerase variant and application thereof

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 58, 61, 94, 96, 119, and 155amino 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&#x2032;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.

Phi29 DNA polymerase and encoding gene and application thereof

Growing community of inventors

Shenzhen, China

Yuliang Dong