Biomics

Вслед за термостабильными ДНК-полимеразами из термофильных эубактерий для проведения ПЦР было обращено внимание на микроорганизмы из другого домена Жизни – на термофильные и гипертермофильные археи, многие из которых способны расти при температурах около 100 °С и выше. С 1991 г. по настоящее время клонировано и секвенировано более трех десятков генов термостабильных ДНК-полимераз термофильных и гипертермофильных архей, представляющих 14 родов и около 30 видов. Наиболее широко представлены ДНК-полимеразы из рода Thermococcus. При этом наиболее часто используемой является Pfu-полимераза из гипертермофильной археи Pyrococcus furiosus. Подавляющее большинство этих ферментов относится к B-семейству ДНК-полимераз и кроме нуклеотидилтрансферазной (полимеразной) активности они имеют еще и редактирующую 3’→5’-экзонуклеазную активность, обеспечивающую увеличенную точность копирования в ПЦР. Большинство этих архейных ДНК-полимераз характеризуются продолжительным временем полужизни при 95 °С и даже при 100 °С, измеряемым для некоторых многими часами. Архейные ДНК-полимеразы нашли применение при амплификации трудных матриц, в том числе протяженных матриц с высоким GC-составом, где себя хорошо проявили смеси таких ферментов с Taq-полимеразой.

археи, гипертермофильные археи, термостабильная ДНК-полимераза, 3`→5`-экзонуклеазная редактирующая активность, ДНК, ПЦР

1. 1. Прангишвили Д.А. ДНК-зависимые ДНК-полимеразы термоацидофильной архебактерии Sulfolobus Молекул. Биология. 1985. 19(2). 477-488.
   2. Слободкина Г.Б., Черных Н.А., Лопатин С.А. и др. Выделение и характеристика термостабильной ДНК-полимеразы гипертермофильной археи Thermococcus litoralis Sh1AM. Прикладн. Биохимия и микробиология. 2005. 41(1). 40-47.
   3. Bae H, Kim KP, Lee JI et al. Characterization of DNA polymerase from the hyperthermophilic archaeon Thermococcus marinus and its application to PCR. Extremophiles. 2009. 13(4). 657-667. doi: 10.1007/s00792-009-0248-0
   4. Böhlke K, Pisani FM, Vorgias CE et al. PCR performance of the B-type DNA polymerase from the thermophilic euryarchaeon Thermococcus aggregans improved by mutations in the Y-GG/A motif. Nucleic Acids Res. 2000. 28(20). 3910-3917. doi: 10.1093/nar/28.20.3910
   5. Cambon-Bonavita MA, Schmitt P, Zieger M et al. Cloning, expression, and characterization of DNA polymerase I from the hyperthermophilic archaea Thermococcus fumicolans. Extremophiles. 2000. 4(4). 215-225. doi: 10.1007/pl00010714
   6. Cann IK, Komori K, Toh H et al. A heterodimeric DNA polymerase: evidence that members of Euryarchaeota possess a distinct DNA polymerase. Proc Natl Acad Sci USA. 1998. 95(24). 14250-14255. doi: 10.1073/pnas.95.24.14250
   7. Ceylan HK. Enhanced Biomass Production of Recombinant Pfu DNA Polymerase Producer Escherichia coli BL21(DE3) by Optimization of Induction Variables Using Response Surface Methodology. Protein J. 42(4). 451-462. doi: 10.1007/s10930-023-10122-8
   8. Chae YK, Jeon W, Cho KS. Rapid and simple method to prepare functional Pfu DNA polymerase expressed in Escherichia coli Journal of Microbiology and Biotechnology. 2002. 12(5). 841-843.
   9. Cho SS, Kim KP, Lee KK et al. Characterization and PCR application of a new high-fidelity DNA polymerase from Thermococcus waiotapuensis. Enzyme Microb Technol. 51(6-7). 334-341. doi: 10.1016/j.enzmictec.2012.07.017
   10. Choi JJ, Nam KH, Min B et al. Protein trans-splicing and characterization of a split family B-type DNA polymerase from the hyperthermophilic archaeal parasite Nanoarchaeum equitans. J Mol Biol. 356(5). 1093-1106. doi: 10.1016/j.jmb.2005.12.036
   11. Choi JJ, Song JG, Nam KH et al. Unique substrate spectrum and PCR application of Nanoarchaeum equitans family B DNA polymerase. Appl Environ Microbiol. 2008. 74(21). 6563-6569. doi: 10.1128/AEM.00624-08
   12. Dabrowski S, Kur J. Cloning and expression in Escherichia coli of the recombinant his-tagged DNA polymerases from Pyrococcus furiosus and Pyrococcus woesei. Protein Expr Purif. 1998. 14(1). 131-138. doi: 10.1006/prep.1998.0945
   13. Daimon K, Ishino S, Imai N et al. Two Family B DNA Polymerases From Aeropyrum pernix, Based on Revised Translational Frames. Front Mol Biosci. 2018. 5. 37. doi: 10.3389/fmolb.2018.00037
   14. Datukishvili N, Pokholok D, Lottspeich F et al. The DNA polymerase-encoding gene from a thermoacidophilic archaeon Sulfolobus acidocaldarius. Gene. 1996. 177(1-2). 271-273. doi: 10.1016/0378-1119(96)00298-3
   15. De Felice M, Medagli B, Esposito L et al. Biochemical evidence of a physical interaction between Sulfolobus solfataricus B-family and Y-family DNA polymerases. Extremophiles. 2007. 11(2). 277-282. doi: 10.1007/s00792-006-0038-x
   16. De Felice M, Sensen CW, Charlebois RL et al. Two DNA polymerase sliding clamps from the thermophilic archaeon Sulfolobus solfataricus. J Mol Biol. 1999. 291(1). 47-57. doi: 10.1006/jmbi.1999.2939
   17. Dietrich J, Schmitt P, Zieger M et al. PCR performance of the highly thermostable proof-reading B-type DNA polymerase from Pyrococcus abyssi. FEMS Microbiol Lett. 2002. 217(1). 89-94. doi: 10.1111/j.1574-6968.2002.tb11460.x
   18. Elie C, De Recondo AM, Forterre P. Thermostable DNA polymerase from the archaebacterium Sulfolobus acidocaldarius. Purification, characterization and immunological properties. Eur J Biochem. 1989. 178(3). 619-626. doi: 10.1111/j.1432-1033.1989.tb14490.x
   19. Elie C, Salhi S, Rossignol JM et al. A DNA polymerase from a thermoacidophilic archaebacterium: evolutionary and technological interests. Biochim Biophys Acta. 1988. 951(2-3). 261-267. doi: 10.1016/0167-4781(88)90095-4
   20. Feng X, Liu X, Xu R et al. A Unique B-Family DNA Polymerase Facilitating Error-Prone DNA Damage Tolerance in Crenarchaeota. Front Microbiol. 2020. 11. 1585. doi: 10.3389/fmicb.2020.01585
   21. Forterre P. The DNA polymerase from the archaebacterium Pyrococcus furiosus does not testify for a specific relationship between archaebacteria and eukaryotes. Nucleic Acids Res. 1992. 20(7). 1811. doi: 10.1093/nar/20.7.1811
   22. Forterre P, Elie C, Sioud M et al. Studies on DNA polymerases and topoisomerases in archaebacteria. Can J Microbiol. 35(1). 228-233. doi: 10.1139/m89-035
   23. Ghasemi A, Salmanian AH, Sadeghifard N et al. Cloning, expression and purification of Pwo polymerase from Pyrococcus woesei. Iran J Microbiol. 2011. 3(3). 118-122.
   24. Goldman S, Kim R, Hung LW et al. Purification, crystallization and preliminary X-ray crystallographic analysis of Pyrococcus furiosus DNA polymerase. Acta Crystallogr D Biol Crystallogr. 1998. 54(5). 986-988. doi: 10.1107/s0907444998000353
   25. Griffiths K, Nayak S, Park K et al. New high fidelity polymerases from Thermococcus Protein Expr Purif. 2007. 52(1). 19-30. doi: 10.1016/j.pep.2006.07.022
   26. Gueguen Y, Rolland JL, Lecompte O et al. Characterization of two DNA polymerases from the hyperthermophilic euryarchaeon Pyrococcus abyssi. Eur J Biochem. 268(22). 5961-5969. doi: 10.1046/j.0014-2956.2001.02550.x
   27. Guo J, Zhang W, Coker AR et al. Structure of the family B DNA polymerase from the hyperthermophilic archaeon Pyrobaculum calidifontis. Acta Crystallogr D Struct Biol. 73(5). 420-427. doi: 10.1107/S2059798317004090
   28. Hadi MI, Laksmi FA, Helbert et al. An efficient approach for overproduction of DNA polymerase from Pyrococcus furiosus using an optimized autoinduction system in Escherichia coli. World J Microbiol Biotechnol. 40(10). 324. doi: 10.1007/s11274-024-04127-3
   29. Hamal A, Forterre P, Elie C. Purification and characterization of a DNA polymerase from the archaebacterium Thermoplasma acido Eur J Biochem. 1990. 190(3). 517-521. doi: 10.1111/j.1432-1033.1990.tb15604.x
   30. Hashimoto H, Matsumoto T, Nishioka M et al. Crystallographic studies on a family B DNA polymerase from hyperthermophilic archaeon Pyrococcus kodakaraensis strain KOD1. J Biochem. 1999. 125(6). 983-986. doi: 10.1093/oxfordjournals.jbchem.a022405
   31. Hashimoto H, Nishioka M, Fujiwara S et al. Crystal structure of DNA polymerase from hyperthermophilic archaeon Pyrococcus kodakaraensis J Mol Biol. 2001. 306(3). 469-477. doi: 10.1006/jmbi.2000.4403
   32. Hodges RA, Perler FB, Noren CJ et al. Protein splicing removes intervening sequences in an archaea DNA polymerase. Nucleic Acids Res. 1992. 20(23). 6153-6157. doi: 10.1093/nar/20.23.6153
   33. Hopfner KP, Eichinger A, Engh RA et al. Crystal structure of a thermostable type B DNA polymerase from Thermococcus gorgonarius. Proc Natl Acad Sci USA. 1999. 96(7). 3600-3605. doi: 10.1073/pnas.96.7.3600
   34. Hu JH, Wang F, Liu CZ. Development of an efficient process intensification strategy for enhancing Pfu DNA polymerase production in recombinant Escherichia coli. Bioprocess Biosyst Eng. 2015. 38(4). 651-659. doi: 10.1007/s00449-014-1304-4
   35. Imamura M, Uemori T, Kato I et al. A non-alpha-like DNA polymerase from the hyperthermophilic archaeon Pyrococcus furiosus. Biol Pharm Bull. 1995. 18(12). 1647-1652. doi: 10.1248/bpb.18.1647
   36. Ishino S, Ishino Y. Comprehensive search for DNA polymerase in the hyperthermophilic archaeon, Pyrococcus Nucleosides Nucleotides Nucleic Acids. 2006. 25(4-6). 681-691. doi: 10.1080/15257770600686485
   37. Ishino Y, Ishino S. DNA polymerases from euryarchaeota. Methods Enzymol. 2001. 334. 249-260. doi: 10.1016/s0076-6879(01)34473-7
   38. Ishino Y, Komori K, Cann IK et al. A novel DNA polymerase family found in Archaea. J Bacteriol. 1998. 180(8). 2232-2236. doi: 10.1128/JB.180.8.2232-2236.1998
   39. Kähler M, Antranikian G. Cloning and characterization of a family B DNA polymerase from the hyperthermophilic crenarchaeon Pyrobaculum islandicum. J Bacteriol. 2000. 182(3). 655-663. doi: 10.1128/JB.182.3.655-663.2000
   40. Khaerunnisa, Laksmi FA, Rahayuningsih M et al. One-step rapid production of DNA polymerase from Pyrococcus furiosus in Escherichia coli system under optimized culture conditions. Prep Biochem Biotechnol. 55(8). 1066-1074. doi: 10.1080/10826068.2025.2483238
   41. Kim KP, Bae H, Kim IH et al. Cloning, expression, and PCR application of DNA polymerase from the hyperthermophilic archaeon, Thermococcus celer. Biotechnol Lett. 33(2). 339-346. doi: 10.1007/s10529-010-0434-2
   42. Kim SW, Kim DU, Kim JK et al. Crystal structure of Pfu, the high fidelity DNA polymerase from Pyrococcus furiosus. Int J Biol Macromol. 2008. 42(4). 356-361. doi: 10.1016/j.ijbiomac.2008.01.010
   43. Kim YJ, Lee HS, Bae SS et al. Cloning, purification, and characterization of a new DNA polymerase from a hyperthermophilic archaeon, Thermococcus NA1. J Microbiol Biotechnol. 2007. 17(7). 1090-1097.
   44. Klimczak LJ, Grummt F, Burger KJ. Purification and characterization of DNA polymerase from the archaebacterium Sulfolobus acid Nucleic Acids Res. 1985. 13(14). 5269-5282. doi: 10.1093/nar/13.14.5269
   45. Klimczak LJ, Grummt F, Burger KJ. Purification and characterization of DNA polymerase from the archaebacterium Methanobacterium thermoautotrophicum. Biochemistry. 25(17). 4850-4855. doi: 10.1021/bi00365a019
   46. Komori K, Ishino Y. Functional interdependence of DNA polymerizing and 3`-->5` exonucleolytic activities in Pyrococcus furiosus DNA polymerase I. Protein Eng. 2000. 13(1). 41-47. doi: 10.1093/protein/13.1.41
   47. Kong H, Kucera RB, Jack WE. Characterization of a DNA polymerase from the hyperthermophile archaea Thermococcus litoralis. Vent DNA polymerase, steady state kinetics, thermal stability, processivity, strand displacement, and exonuclease activities. J Biol Chem. 1993. 268(3). 1965-1975. doi: 10.1016/S0021-9258(18)53949-1
   48. Konisky J, Paule SM, Carinato ME et al. The DNA polymerase gene from the methanogenic archaeon Methanococcus voltae. J Bacteriol. 176(20). 6402-6403. doi: 10.1128/jb.176.20.6402-6403.1994
   49. Koonin EV, Krupovic M, Ishino S eet al. The replication machinery of LUCA: common origin of DNA replication and transcription. BMC Biol. 2020. 18(1). 61. doi: 10.1186/s12915-020-00800-9
   50. Kuroita T, Matsumura H, Yokota N et al. Structural mechanism for coordination of proofreading and polymerase activities in archaeal DNA polymerases. J Mol Biol. 2005. 351(2). 291-298. doi: 10.1016/j.jmb.2005.06.015
   51. Kuznetsova AA, Soloveva MA, Mikushina ES et al. Characterization and PCR Application of Family B DNA Polymerases from Thermococcus stetteri. Life (Basel). 14(12). 1544. doi: 10.3390/life14121544
   52. Kwon KM, Kang SG, Sokolova TG et al. Characterization of a family B DNA polymerase from Thermococcus barophilus Ch5 and its application for long and accurate PCR. Enzyme Microb Technol. 2016. 86. 117-1 doi: 10.1016/j.enzmictec.2016.02.004
   53. Lee JI, Cho SS, Kil E-J et al. Characterization and PCR application of a thermostable DNA polymerase from Thermococcus pacificus. Enzyme and Microbial Technology. 2010. 47(4). 147-152. doi: 10.1016/j.enzmictec.2010.06.003
   54. Lee JI, Kim YJ, Bae H et al. Biochemical properties and PCR performance of a family B DNA polymerase from hyperthermophilic Euryarchaeon Thermococcus peptonophilus. Appl Biochem Biotechnol. 160(6). 1585-1599. doi: 10.1007/s12010-009-8658-0
   55. Lee Y-J, Choi JJ, Kwon S-T. Cloning, expression, and partial characterization of a family B-type DNA polymerase from the hyperthermophilic crenarchaeon Sulfophobococcus zilligii. Enzyme and Microbial Technology. 2006. 38(6). 821-830. doi: 10.1016/j.enzmictec.2005.08.010
   56. Li B, Gu C, Zhao J. Molecular cloning and expression of Pfu polymeras gen and its application in long-distanc PCR. Chinese Science Bulletin. 1998. 43(10). 863-867.
   57. Loan TD, Easton CJ, Alissandratos A. DNA amplification with in situ nucleoside to dNTP synthesis, using a single recombinant cell lysate of coli. Sci Rep. 2019. 9(1). 15621. doi: 10.1038/s41598-019-51917-z
   58. Lu C, Erickson HP. Expression in Escherichia coli of the thermostable DNA polymerase from Pyrococcus furiosus. Protein Expr Purif. 11(2). 179-184. doi: 10.1006/prep.1997.0775
   59. Lundberg KS, Shoemaker DD, Adams MW et al. High-fidelity amplification using a thermostable DNA polymerase isolated from Pyrococcus furiosus. Gene. 1991. 108(1). 1-6. doi: 10.1016/0378-1119(91)90480-y
   60. Makarova KS, Krupovic M, Koonin EV. Evolution of replicative DNA polymerases in archaea and their contributions to the eukaryotic replication machinery. Front Microbiol. 2014. 5. doi: 10.3389/fmicb.2014.00354
   61. Marsic D, Flaman JM, Ng JD. New DNA polymerase from the hyperthermophilic marine archaeon Thermococcus thioreducens. Extremophiles. 2008. 12(6). 775-788. doi: 10.1007/s00792-008-0181-7
   62. Mathur EJ, Adams MW, Callen WN et al. The DNA polymerase gene from the hyperthermophilic marine archaebacterium, Pyrococcus furiosus, shows sequence homology with alpha-like DNA polymerases. Nucleic Acids Res. 1991. 19(24). 6952. doi: 10.1093/nar/19.24.6952
   63. Mattila P, Korpela J, Tenkanen T et al. Fidelity of DNA synthesis by the Thermococcus litoralis DNA polymerase - an extremely heat stable enzyme with proofreading activity. Nucleic Acids Res. 19(18). 4967-4973. doi: 10.1093/nar/19.18.4967
   64. Melissis S, Labrou NE, Clonis YD. Nucleotide-mimetic synthetic ligands for DNA-recognizing enzymes One-step purification of Pfu DNA polymerase. J Chromatogr A. 1122(1-2). 63-75. doi: 10.1016/j.chroma.2006.04.044
   65. Mroczkowski BS, Huvar A, Lernhardt W et al. Secretion of thermostable DNA polymerase using a novel baculovirus vector. J Biol Chem. 1994. 269(18). 13522-13528. doi: 10.1016/S0021-9258(17)36862-X
   66. Nastopoulos V, Pisani FM, Savino C et al. Crystallization and preliminary X-ray diffraction studies of DNA polymerase from the thermophilic archaeon Sulfolobus solfataricus. Acta Crystallogr D Biol Crystallogr. 1998. 54(5). 1002-1004. doi: 10.1107/s0907444998002443
   67. Niehaus F, Frey B, Antranikian G. Cloning and characterisation of a thermostable alpha-DNA polymerase from the hyperthermophilic archaeon Thermococcus TY. Gene. 1997. 204(1-2). 153-158. doi: 10.1016/s0378-1119(97)00536-2
   68. Nishida H, Tanabe M, Ishino Y Crystallization and preliminary crystallographic study of DNA polymerase from Pyrococcus furiosus. Protein Pept Lett. 2007. 14(4). 403-405. doi: 10.2174/092986607780363853
   69. Nuryana I, Laksmi FA, Dewi KS, et al. Codon optimization of a gene encoding DNA polymerase from Pyrococcus furiosus and its expression in Escherichia coli. J Genet Eng Biotechnol. 21(1). 129. doi: 10.1186/s43141-023-00605-7
   70. Perler FB, Comb DG, Jack WE et al. Intervening sequences in an Archaea DNA polymerase gene. Proc Natl Acad Sci USA. 1992. 89(12). 5577-5581. doi: 10.1073/pnas.89.12.5577
   71. Pisani FM, De Martino C, Rossi M. A DNA polymerase from the archaeon Sulfolobus solfataricus shows sequence similarity to family B DNA polymerases. Nucleic Acids Res. 1992. 20(11). 2711-2716. doi: 10.1093/nar/20.11.2711
   72. Pisani FM, De Felice M, Rossi M. Amino acid residues involved in determining the processivity of the 3`-5` exonuclease activity in a family B DNA polymerase from the thermoacidophilic archaeon Sulfolobus solfataricus. Biochemistry. 37(42). 15005-15012. doi: 10.1021/bi981127s
   73. Pisani FM, Manco G, Carratore V et al. Domain organization and DNA-induced conformational changes of an archaeal family B DNA polymerase. Biochemistry. 1996. 35(28). 9158-9166. doi: 10.1021/bi960446l
   74. Pisani FM, Rossi M. Evidence that an archaeal alpha-like DNA polymerase has a modular organization of its associated catalytic J Biol Chem. 1994. 269(11). 7887-7892. doi: 10.1016/S0021-9258(17)37134-X
   75. Prangishvili D, Klenk HP. Nucleotide sequence of the gene for a 74 kDa DNA polymerase from the archaeon Sulfolobus solfataricu Nucleic Acids Res. 1993. 21(11). 2768. doi: 10.1093/nar/21.11.2768
   76. Raffin JP, Henneke G, Dietrich J. Purification and characterization of a new DNA polymerase modulator from the hyperthermophilic archaeon Thermococcus fumicolans. Comp Biochem Physiol B Biochem Mol Biol. 2000. 127(3). 299-308. doi: 10.1016/s0305-0491(00)00263-7
   77. Rella R, Raia CA, Pisani FM et al. Purification and properties of a thermophilic and thermostable DNA polymerase from the archaebacterium Sulfolobus solfataricus. Ital J Biochem. 39(2). 83-99.
   78. Rogozin IB, Makarova KS, Pavlov YI et al. A highly conserved family of inactivated archaeal B family DNA polymerases. Biol Direct. 3. 32. doi: 10.1186/1745-6150-3-32
   79. Rodriguez AC, Park HW, Mao C et al. Crystal structure of a pol alpha family DNA polymerase from the hyperthermophilic archaeon Thermococcus 9 degrees N-7. J Mol Biol. 2000. 299(2). 447-462. doi: 10.1006/jmbi.2000.3728
   80. Rossi M, Rella R, Pensa M et al. Structure and properties of a thermophylic and thermostable DNA polymerase isolated from Sulfolobus solfataricus. System. Appl. Microbil. 1986. 7. 337-341. doi: 10.1016/S0723-2020(86)80029-7
   81. Salhi S, Elie C, Forterre P et al. DNA polymerase from Sulfolobus acidocaldarius. Replication at high temperature of long stretches of single-stranded DNA. J Mol Biol. 1989. 209(4). 635-644. doi: 10.1016/0022-2836(89)91000-0
   82. Sankar PS, Citartan M, Siti AA et al. A simple method for in-house Pfu DNA polymerase purification for high-fidelity PCR amplification. Iran J Microbiol. 2019. 11(2). 181-186.
   83. Sarmiento F, Long F, Cann I et al. Diversity of the DNA Replication System in the Archaea Domain. 2014. 2014. 675946. doi: 10.1155/2014/675946
   84. Savino C, Federici L, Johnson KA et al. Insights into DNA replication: the crystal structure of DNA polymerase B1 from the archaeon Sulfolobus solfataricus. Structure. 2004. 12(11). 2001-2008. doi: 10.1016/j.str.2004.09.007
   85. Segerer AH, Burggraf S, Fiala G et al. Life in hot springs and hydrothermal vents. Orig Life Evol Biosph. 23(1). 77-90. doi: 10.1007/BF01581992
   86. Seo KJ, Cho SS, Ppyun HW et al. Characterization of a family B DNA polymerase from the hyperthermophilic crenarchaeon Ignicoccus hospitalis KIN4/I and its application to PCR. Appl Biochem Biotechnol. 173(5). 1108-1120. doi: 10.1007/s12010-014-0918-y
   87. Stetter KO. History of discovery of the first hyperthermophiles. Extremophiles. 2006. 10(5). 357-362. doi: 10.1007/s00792-006-0012-7
   88. Southworth MW, Kong H, Kucera RB et al. Cloning of thermostable DNA polymerases from hyperthermophilic marine Archaea with emphasis on Thermococcus 9 degrees N-7 and mutations affecting 3`-5` exonuclease activity. Proc Natl Acad Sci USA. 1996. 93(11). 5281-5285. doi: 10.1073/pnas.93.11.5281
   89. Sun Z, Cai J. Purification of recombinant Pfu DNA polymerase using a new JK110 resin. Korean J. Chem. Eng. 2006. 23. 607-609.
   90. Takagi M, Nishioka M, Kakihara H et al. Characterization of DNA polymerase from Pyrococcus strain KOD1 and its application to PCR. Appl Environ Microbiol. 1997. 63(11). 4504-4510. doi: 10.1128/aem.63.11.4504-4510.1997
   91. Terpe K. Overview of thermostable DNA polymerases for classical PCR applications: from molecular and biochemical fundamentals to commercial systems. Appl Microbiol Biotechnol. 2013. 97(24). 10243-10254. doi: 10.1007/s00253-013-5290-2
   92. Uemori T, Ishino Y, Doi H et al. The hyperthermophilic archaeon Pyrodictium occultum has two alpha-like DNA polymerases. J Bacteriol. 177(8). 2164-2177. doi: 10.1128/jb.177.8.2164-2177.1995
   93. Uemori T, Ishino Y, Toh H et al. Organization and nucleotide sequence of the DNA polymerase gene from the archaeon Pyrococcus furiosus. Nucleic Acids Res. 1993. 21(2). 259-265. doi: 10.1093/nar/21.2.259
   94. Uemori T, Sato Y, Kato I et al. A novel DNA polymerase in the hyperthermophilic archaeon, Pyrococcus furiosus: gene cloning, expression, and characterization. Genes Cells. 1997. 2(8). 499-512. doi: 10.1046/j.1365-2443.1997.1380336.x
   95. Wang F, Li S, Zhao H et al. Expression and Characterization of the RKOD DNA Polymerase in Pichia pastoris. PLoS One. 2015. 10(7). e0131757. doi: 10.1371/journal.pone.0131757
   96. Yamashita M, Xu J, Morokuma D et al. Characterization of Recombinant Thermococcus kodakaraensis (KOD) DNA Polymerases Produced Using Silkworm-Baculovirus Expression Vector System. Mol Biotechnol. 59(6). 221-233. doi: 10.1007/s12033-017-0008-9
   97. Zhang L, Jiang D, Shi H et al. Characterization and application of a family B DNA polymerase from the hyperthermophilic and radioresistant euryarchaeon Thermococcus gammatolerans. Int J Biol Macromol. 156. 217-224. doi: 10.1016/j.ijbiomac.2020.03.204
   98. Zheng W, Wang Q, Bi Q. Construction, Expression, and Characterization of Recombinant Pfu DNA Polymerase in Escherichia coli. Protein J. 35(2). 145-153. doi: 10.1007/s10930-016-9651-4
   99. Zhao Y, Jeruzalmi D, Moarefi I et al. Crystal structure of an archaebacterial DNA polymerase. Structure. 1999. 7(10). 1189-1199. doi: 10.1016/s0969-2126(00)80053-2
   100. Zhou M, Mao C, Rodriguez AC et al. Crystallization and preliminary diffraction analysis of a hyperthermostable DNA polymerase from a Thermococcus archaeon. Acta Crystallogr D Biol Crystallogr. 1998. 54(5). 994-995. doi: 10.1107/s0907444998001553