公开(公告)号：EP4435098A1

公开(公告)日：2024-09-25

申请号：EP21963712.1

申请日：2021-11-15

申请人： BGI Shenzhen

发明人： AN, Qun ,  LIN, Qibin ,  GUO, Fei ,  XIE, Qingqing ,  ZHENG, Yue ,  XI, Feng ,  DONG, Yuliang ,  ZHANG, Wenwei

IPC分类号： C12N9/12 ,  C12N15/54 ,  C12P19/34

CPC分类号： C12P19/34 ,  C12N9/10 ,  C12N9/12

摘要： Provided is a chimeric DNA polymerase, comprising: a first peptide segment, which has at least 80% homology with at least part of the amino acid sequence in the N-terminal domain of a 90N DNA polymerase; a second peptide segment, which has at least 80% homology with at least part of the amino acid sequence in the exonucleolytic domain of a KOD DNA polymerase; a third peptide segment, which has at least 80% homology with at least some of the amino acids in the N-terminal domain of the 90N DNA polymerase; a fourth peptide segment, which has at least 80% homology with at least some of the amino acids in the palm domain of a KOD DNA polymerase; a fifth peptide segment, which has at least 80% homology with at least some of the amino acids in the finger domain of the Pfu DNA polymerase; a sixth peptide segment, which has at least 80% homology with at least some of the amino acids in the palm domain of the KOD DNA polymerase; and a seventh peptide segment, which has at least 80% homology with at least some of the amino acids in the thumb domain of the 90N DNA polymerase.

公开(公告)号：EP4458958A1

公开(公告)日：2024-11-06

申请号：EP21969852.9

申请日：2021-12-31

申请人： BGI Shenzhen

发明人： ZHANG, Wei ,  DONG, Yuliang ,  LI, Jing ,  ZHENG, Yue ,  ZHANG, Wenwei

IPC分类号： C12N9/02 ,  C12Q1/66

摘要： A mutant and the uses thereof. Compared with the amino acid sequence shown in SEQ ID NO: 2, the mutant has at least one of the following mutation sites: the 24th, 26th, 27th, 29th, 30th, 31st, 32nd, 33rd, 36th, 37th, 40th, 66th, 79th, 84th, 88th, 102nd, 103rd, 104th, 110th, 123rd, 124th, 138th, 152nd, 163rd, 167th, 170th, 174th, 175th, 178th, 182nd and 183rd sites.

公开(公告)号：EP4455279A1

公开(公告)日：2024-10-30

申请号：EP21968652.4

申请日：2021-12-24

申请人： BGI Shenzhen

发明人： ZHANG, Xiaohong ,  LAN, Xi ,  XIE, Qingqing ,  XI, Feng ,  ZHENG, Yue ,  DONG, Yuliang ,  ZHANG, Wenwei

IPC分类号： C12N9/12 ,  C12N15/54 ,  C12Q1/686

摘要： The present invention relates to the field of biotechnology, in particular to a DNA polymerase mutant and the use thereof. Compared with a wild type, the mutant provided in the present invention has a significantly improved DNA polymerization activity, significantly improved affinity to DNA and amplification uniformity; and has good effects in the amplification of multiplex PCR, high-GC templates and complex templates. A hot-start-version DNA polymerase is further prepared by means of using antibodies or chemical modifications, wherein the yield of the DNA polymerase is significantly improved, while the primer dimer is significantly decreased. The prepared mutant hot-start DNA polymerase can be applied to PCR amplification of multiplex PCR high-GC templates and complex templates.

公开(公告)号：EP3854872A1

公开(公告)日：2021-07-28

申请号：EP18936847.5

申请日：2018-10-11

申请人： BGI Shenzhen

发明人： ZHANG, Zhougang ,  LIU, Huanhuan ,  ZHENG, Yue ,  ZHOU, Yujun ,  XIA, Jun ,  DONG, Yuliang ,  XU, Chongjun ,  ZHANG, Wenwei

IPC分类号： C12N9/12 ,  C12N15/52 ,  C12N15/63

摘要： 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).

公开(公告)号：EP4234690A1

公开(公告)日：2023-08-30

申请号：EP20958065.3

申请日：2020-10-21

申请人： BGI Shenzhen

发明人： ZHAI, Lili ,  XIE, Qingqing ,  CAO, Ruyin ,  WANG, Zi ,  ZHENG, Yue ,  DONG, Yuliang ,  ZHANG, Wenwei ,  XU, Xun

IPC分类号： C12N9/12 ,  C12Q1/68

摘要： Provided is a recombinant KOD polymerase. A KOD polymerase mutant is a protein as follows: the protein is a protein having DNA polymerase activity that is obtained by modifying amino acid residues in at least one of the following 48 positions in the amino acid sequence of KOD DNA polymerase GH78: 267, 326, 347, 349, 353, 375, 378, 379, 380, 385, 451, 452, 453, 454, 457, 461, 465, 470, 474, 477, 478, 479, 480, 482, 484, 485, 486, 493, 496, 497, 514, 574, 584, 605, 610, 630, 665, 666, 667, 674, 676, 680, 682, 698, 707, 718, 723 and 729, without changing other amino acid sequences; and compared with KOD DNA polymerase GH78, with regard to catalysis, the recombinant DNA polymerase exhibits a faster reaction rate, better catalytic efficiency, better affinity and other advantages, thereby improving the reaction rate of DNA polymerase in sequencing and increasing the reaction read length.

公开(公告)号：EP4349977A1

公开(公告)日：2024-04-10

申请号：EP21942622.8

申请日：2021-09-08

申请人： BGI Shenzhen

发明人： ZHAI, Lili ,  XIE, Qingqing ,  CAO, Ruyin ,  ZHENG, Yue ,  DONG, Yuliang ,  ZHANG, Wenwei ,  XU, Xun ,  CHEN, Zhuo

IPC分类号： C12N9/12 ,  C12Q1/68

摘要： Provided are a thermostable B-family DNA polymerase mutant and an application thereof. The thermostable B-family DNA polymerase mutant is provided to mutate amino acid residues in at least two of three sites of group A, i.e., 408, 409, and 410 in an amino acid sequence of thermostable B-family DNA polymerases to obtain proteins having DNA polymerase activity. A-group sites (408/409/410), B-group sites (451/485), C-group sites (389/383/384), D-group sites (589/67/680), E-group sites (491/493/494/497), and F-group sites (474/478/486/480/484) of several B-family DNA polymerases are analyzed to design the mutant. Compared with non-mutant DNA polymerases, several groups of mutant DNA polymerases exhibit a strong polymerization capability in catalyzing 3' blocking-modified nucleotides and provide a basis for the sustainable development of high-throughput sequencing.

公开(公告)号：EP4166659A1

公开(公告)日：2023-04-19

申请号：EP20939885.8

申请日：2020-10-10

申请人： BGI Shenzhen

发明人： ZHENG, Yue ,  XU, Xun ,  ZHANG, Wenwei ,  DONG, Yuliang ,  ZHOU, Yujun

IPC分类号： C12N9/12 ,  C12N15/54 ,  C12N15/70 ,  C12P19/34 ,  C12Q1/68

摘要： Provided are a stable Phi29 DNA polymerase having high enzyme activity, and an encoding gene and an application thereof. The Phi29 DNA polymerase is obtained by substituting amino acid residue at at least one among the following 29 positions: position 17, position 96, position 97, position 99, position 123, position 140, position 148, position 158, position 159, position 171, position 203, position 204, position 213, position 217, position 224, position 250, position 270, position 309, position 310, position 320, position 344, position 345, position 347, position 369, position 402, position 416, position 509, position 515 and position 524 of a DNA polymerase shown in SEQ ID NO: 2.

公开(公告)号：EP3862427A1

公开(公告)日：2021-08-11

申请号：EP18932681.2

申请日：2018-09-03

申请人： BGI Shenzhen

发明人： ZHAI, Lili ,  WANG, Lin ,  ZHANG, Wenwei ,  DONG, Yuliang ,  ZHENG, Yue ,  LIU, Fen

IPC分类号： C12N9/12 ,  C12N15/52 ,  C12N15/63

摘要： 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).