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A PK15 cell line stably expressing prolyl oligopeptidase (POP) and its effect on the proliferation of foot-and-mouth disease virus
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Ziyi WANG1, 2, 3, Yi RU1, 2, Bingzhou LU1, 2, Yang YANG1, 2, Longhe ZHAO1, 2, Yajun LI1, 2, Jianbin LI1, 2, Minggui LI1, 2, Kun MA1, 2, Feifan LENG3, Rongzeng HAO1, 2, Haixue ZHENG1, 2, 3
Acta Microbiologica Sinica | 2025, 65(10) : 4458 - 4471
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Acta Microbiologica Sinica | 2025, 65(10): 4458-4471
• Research Article •
A PK15 cell line stably expressing prolyl oligopeptidase (POP) and its effect on the proliferation of foot-and-mouth disease virus
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Ziyi WANG1, 2, 3, Yi RU1, 2, Bingzhou LU1, 2, Yang YANG1, 2, Longhe ZHAO1, 2, Yajun LI1, 2, Jianbin LI1, 2, Minggui LI1, 2, Kun MA1, 2, Feifan LENG3, Rongzeng HAO1, 2, Haixue ZHENG1, 2, 3
Affiliations
  • 1 State Key Laboratory for Animal Disease Control and Prevention, College of Veterinary Medicine of Lanzhou University, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu, China
  • 2 Gansu Province Research Center for Basic Discipline of Pathogen Biology, Lanzhou, Gansu, China
  • 3 School of Life Science and Engineering, Lanzhou University of Technology, Lanzhou, Gansu, China
Published: 2025-09-04 doi: 10.13343/j.cnki.wsxb.20250191
Outline
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[Objective] To construct a PK15 cell line stably expressing the prolyl oligopeptidase (POP) by using the PiggyBac transposon system and systematically investigate its regulatory role in the proliferation of foot-and-mouth disease virus (FMDV). [Methods] A recombinant PiggyBac vector carrying the POP gene was constructed and transfected into PK15 cells, and the expression of the target protein was detected by Western blotting. The monoclonal cell line stably expressing POP was isolated via the limiting dilution method. The stability of mRNA and protein levels of POP was verified by RT-qPCR and Western blotting, respectively. The viability of the selected cell line was assessed by the Cell Counting Kit-8 (CCK-8) assay. FMDV replication kinetics in the stable cell line were comprehensively analyzed by Western blotting, RT-qPCR, and the 50% tissue culture infective dose (TCID50) assay. [Results] A PK15 cell line stably expressing POP was established. No significant differences in cell viability were observed between the stable cell line and the control cell line. The protein level of POP remained stable in the established cell line after 30 passages. The results of the viral infection assay demonstrated that the FMDV proliferation level in the PK15 cell line stably expressing POP was significantly higher than that in the control group, with this stimulatory effect being maintained across multiple passages. [Conclusion] We successfully constructed a PK15 cell line stably expressing POP. The results reveal that POP overexpression enhances FMDV replication in a passage-independent manner. These findings provide a valuable experimental model for elucidating the molecular mechanism underlying the role of POP in FMDV replication.

prolyl oligopeptidase (POP)  /  PiggyBac transposition system  /  PK15 cells  /  virus proliferation
Ziyi WANG, Yi RU, Bingzhou LU, Yang YANG, Longhe ZHAO, Yajun LI, Jianbin LI, Minggui LI, Kun MA, Feifan LENG, Rongzeng HAO, Haixue ZHENG. A PK15 cell line stably expressing prolyl oligopeptidase (POP) and its effect on the proliferation of foot-and-mouth disease virus[J]. Acta Microbiologica Sinica, 2025 , 65 (10) : 4458 -4471 . DOI: 10.13343/j.cnki.wsxb.20250191
Funding
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  • the National Natural Science Foundation of China(32372990)
  • the Natural Science Foundation of Gansu Province(23YFNA0011)
Appendix
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Year 2025 volume 65 Issue 10
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117
46
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Article Info
doi: 10.13343/j.cnki.wsxb.20250191
  • Receive Date：2025-03-09
  • Online Date：2025-11-03
  • Published：2025-09-04
Article Data
Affiliations
History
  • Received：2025-03-09
  • Accepted：2025-04-25
Funding
the National Natural Science Foundation of China(32372990)
the Natural Science Foundation of Gansu Province(23YFNA0011)
Affiliations
    1 State Key Laboratory for Animal Disease Control and Prevention, College of Veterinary Medicine of Lanzhou University, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu, China
    2 Gansu Province Research Center for Basic Discipline of Pathogen Biology, Lanzhou, Gansu, China
    3 School of Life Science and Engineering, Lanzhou University of Technology, Lanzhou, Gansu, China

Corresponding:

E-mail: ZHENG Haixue,
HAO Rongzeng,
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表12种不同金属材料的力学参数
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Family		 属数
Number of
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Number of
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Percentage of
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鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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