Keeping all secondary structures of the non-coding region in the circular genome of human bocavirus 2 is important for DNA replication and virus assembly, as revealed by three hetero-recombinant genomic clones
The episomal structures of all human bocavirus (HBoV) genotypes have been deciphered, including the circular genome of HBoV2 (HBoV2-C1). To discern the role of the circular HBoV2 genome, three distinct linearized HBoV2-C1 genomes were cloned into pBlueScript SKII(+) to obtain pBlueScript HBoV2 5043–...
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Taylor & Francis Group
2019-01-01
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| Series: | Emerging Microbes and Infections |
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| Online Access: | https://www.tandfonline.com/doi/10.1080/22221751.2019.1682949 |
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| author | Linqing Zhao Tao Wang Yuan Qian Jingdong Song Runan Zhu Liying Liu Liping Jia Huijin Dong |
| author_facet | Linqing Zhao Tao Wang Yuan Qian Jingdong Song Runan Zhu Liying Liu Liping Jia Huijin Dong |
| author_sort | Linqing Zhao |
| collection | DOAJ |
| description | The episomal structures of all human bocavirus (HBoV) genotypes have been deciphered, including the circular genome of HBoV2 (HBoV2-C1). To discern the role of the circular HBoV2 genome, three distinct linearized HBoV2-C1 genomes were cloned into pBlueScript SKII(+) to obtain pBlueScript HBoV2 5043–5042 (retaining all secondary structures), pBlueScript-HBoV2 5075–5074 (retaining hairpin number 2 and the 5′ terminal structure), and pBlueScript-HBoV2 5220–5219 (retaining only the 5′ terminal structure at the 5′ -genome end). The recombinant plasmids were separately transfected HEK293 cells, revealing that more HBoV2 DNA had accumulated in the pBlueScript HBoV2 5043–5042-transfected HEK293 cells at 72 h post-transfection, as determined by real-time PCR. However, more mRNA was transcribed by pBlueScript-HBoV2 5075–5074 than by the other constructs, as determined by dot-blot hybridization and RNAscope. No significant differences in NS1-70 protein expression were observed among the three HBoV2 genomic clones. However, electron microscopy showed that HBoV2 virus particles were only present in the pBlueScript HBoV2 5043–5042-transfected HEK293 cells. By using three hetero-recombinant HBoV2 genomic clones in HEK293 transfected cells, only the genome with intact secondary structures produced virus particles, suggesting that retaining these structures in a circular genome is important for HBoV2 DNA replication and virus assembly. |
| format | Article |
| id | doaj-art-fb9369459bc4489ba86d772d552a1ff6 |
| institution | OA Journals |
| issn | 2222-1751 |
| language | English |
| publishDate | 2019-01-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | Emerging Microbes and Infections |
| spelling | doaj-art-fb9369459bc4489ba86d772d552a1ff62025-08-20T02:26:09ZengTaylor & Francis GroupEmerging Microbes and Infections2222-17512019-01-01811563157310.1080/22221751.2019.1682949Keeping all secondary structures of the non-coding region in the circular genome of human bocavirus 2 is important for DNA replication and virus assembly, as revealed by three hetero-recombinant genomic clonesLinqing Zhao0Tao Wang1Yuan Qian2Jingdong Song3Runan Zhu4Liying Liu5Liping Jia6Huijin Dong7Laboratory of Virology, Beijing Key Laboratory of Etiology of Viral Diseases in Children, Capital Institute of Pediatrics, Beijing, People’s Republic of ChinaLaboratory of Virology, Beijing Key Laboratory of Etiology of Viral Diseases in Children, Capital Institute of Pediatrics, Beijing, People’s Republic of ChinaLaboratory of Virology, Beijing Key Laboratory of Etiology of Viral Diseases in Children, Capital Institute of Pediatrics, Beijing, People’s Republic of ChinaState Key Laboratory for Infectious Disease Prevention and Control, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing, People’s Republic of ChinaLaboratory of Virology, Beijing Key Laboratory of Etiology of Viral Diseases in Children, Capital Institute of Pediatrics, Beijing, People’s Republic of ChinaLaboratory of Virology, Beijing Key Laboratory of Etiology of Viral Diseases in Children, Capital Institute of Pediatrics, Beijing, People’s Republic of ChinaLaboratory of Virology, Beijing Key Laboratory of Etiology of Viral Diseases in Children, Capital Institute of Pediatrics, Beijing, People’s Republic of ChinaLaboratory of Virology, Beijing Key Laboratory of Etiology of Viral Diseases in Children, Capital Institute of Pediatrics, Beijing, People’s Republic of ChinaThe episomal structures of all human bocavirus (HBoV) genotypes have been deciphered, including the circular genome of HBoV2 (HBoV2-C1). To discern the role of the circular HBoV2 genome, three distinct linearized HBoV2-C1 genomes were cloned into pBlueScript SKII(+) to obtain pBlueScript HBoV2 5043–5042 (retaining all secondary structures), pBlueScript-HBoV2 5075–5074 (retaining hairpin number 2 and the 5′ terminal structure), and pBlueScript-HBoV2 5220–5219 (retaining only the 5′ terminal structure at the 5′ -genome end). The recombinant plasmids were separately transfected HEK293 cells, revealing that more HBoV2 DNA had accumulated in the pBlueScript HBoV2 5043–5042-transfected HEK293 cells at 72 h post-transfection, as determined by real-time PCR. However, more mRNA was transcribed by pBlueScript-HBoV2 5075–5074 than by the other constructs, as determined by dot-blot hybridization and RNAscope. No significant differences in NS1-70 protein expression were observed among the three HBoV2 genomic clones. However, electron microscopy showed that HBoV2 virus particles were only present in the pBlueScript HBoV2 5043–5042-transfected HEK293 cells. By using three hetero-recombinant HBoV2 genomic clones in HEK293 transfected cells, only the genome with intact secondary structures produced virus particles, suggesting that retaining these structures in a circular genome is important for HBoV2 DNA replication and virus assembly.https://www.tandfonline.com/doi/10.1080/22221751.2019.1682949Human bocavirus 2circular genomegenomic clonesDNA replicationvirus assembly |
| spellingShingle | Linqing Zhao Tao Wang Yuan Qian Jingdong Song Runan Zhu Liying Liu Liping Jia Huijin Dong Keeping all secondary structures of the non-coding region in the circular genome of human bocavirus 2 is important for DNA replication and virus assembly, as revealed by three hetero-recombinant genomic clones Emerging Microbes and Infections Human bocavirus 2 circular genome genomic clones DNA replication virus assembly |
| title | Keeping all secondary structures of the non-coding region in the circular genome of human bocavirus 2 is important for DNA replication and virus assembly, as revealed by three hetero-recombinant genomic clones |
| title_full | Keeping all secondary structures of the non-coding region in the circular genome of human bocavirus 2 is important for DNA replication and virus assembly, as revealed by three hetero-recombinant genomic clones |
| title_fullStr | Keeping all secondary structures of the non-coding region in the circular genome of human bocavirus 2 is important for DNA replication and virus assembly, as revealed by three hetero-recombinant genomic clones |
| title_full_unstemmed | Keeping all secondary structures of the non-coding region in the circular genome of human bocavirus 2 is important for DNA replication and virus assembly, as revealed by three hetero-recombinant genomic clones |
| title_short | Keeping all secondary structures of the non-coding region in the circular genome of human bocavirus 2 is important for DNA replication and virus assembly, as revealed by three hetero-recombinant genomic clones |
| title_sort | keeping all secondary structures of the non coding region in the circular genome of human bocavirus 2 is important for dna replication and virus assembly as revealed by three hetero recombinant genomic clones |
| topic | Human bocavirus 2 circular genome genomic clones DNA replication virus assembly |
| url | https://www.tandfonline.com/doi/10.1080/22221751.2019.1682949 |
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