Replication of norovirus in cultured cells
Human Norovirus (NoV), a positive-sense RNA virus, is well-known for causing epidemic gastroenteritis worldwide. Over the past decades, studies on NoV have been restricted due to a lack of appropriate cell culture system. Based on the hypothesis that NoV could be propagated by circumventing the vira...
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| Format: | Final Year Project (FYP) |
| Language: | English |
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2014
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| Online Access: | http://hdl.handle.net/10356/59590 |
| _version_ | 1811677491660587008 |
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| author | Chia, Pei Shan |
| author2 | Liu Ding Xiang |
| author_facet | Liu Ding Xiang Chia, Pei Shan |
| author_sort | Chia, Pei Shan |
| collection | NTU |
| description | Human Norovirus (NoV), a positive-sense RNA virus, is well-known for causing epidemic gastroenteritis worldwide. Over the past decades, studies on NoV have been restricted due to a lack of appropriate cell culture system. Based on the hypothesis that NoV could be propagated by circumventing the viral entry step, this study aims to deliver the naked NoV genome into different cell lines, in order to develop a reliable cell culture system. However, in the absence of a full length NoV genome, the construction of the full length cDNA genome was firstly conducted. Molecular cloning was selected as the main approach, which involved steps such asrestriction enzyme digestion, ligation and transformation. As a result, NoV nucleotide 1-5456 fragments were successfully isolated and cloned in CMV/pBR322(-) plasmid vector. However, subsequent attempts in joining NoV nucleotide 5421-7559 fragments with the upstream nucleotides were unsuccessful. Challenges had surfaced from the three-part ligations performed, which could lead to various combinations of products, therefore further reducing the cloning efficiency. Besides, the cloned full length genome might be toxic or incompatible to the transformed bacteria, causing deleterious mutation effects or death of cells harboring the genome. In short, this study provides reference for future relevant studies and suggestions for improvement of the strategies employed. |
| first_indexed | 2024-10-01T02:38:13Z |
| format | Final Year Project (FYP) |
| id | ntu-10356/59590 |
| institution | Nanyang Technological University |
| language | English |
| last_indexed | 2024-10-01T02:38:13Z |
| publishDate | 2014 |
| record_format | dspace |
| spelling | ntu-10356/595902023-02-28T18:02:22Z Replication of norovirus in cultured cells Chia, Pei Shan Liu Ding Xiang School of Biological Sciences Huang Mei DRNTU::Science::Biological sciences Human Norovirus (NoV), a positive-sense RNA virus, is well-known for causing epidemic gastroenteritis worldwide. Over the past decades, studies on NoV have been restricted due to a lack of appropriate cell culture system. Based on the hypothesis that NoV could be propagated by circumventing the viral entry step, this study aims to deliver the naked NoV genome into different cell lines, in order to develop a reliable cell culture system. However, in the absence of a full length NoV genome, the construction of the full length cDNA genome was firstly conducted. Molecular cloning was selected as the main approach, which involved steps such asrestriction enzyme digestion, ligation and transformation. As a result, NoV nucleotide 1-5456 fragments were successfully isolated and cloned in CMV/pBR322(-) plasmid vector. However, subsequent attempts in joining NoV nucleotide 5421-7559 fragments with the upstream nucleotides were unsuccessful. Challenges had surfaced from the three-part ligations performed, which could lead to various combinations of products, therefore further reducing the cloning efficiency. Besides, the cloned full length genome might be toxic or incompatible to the transformed bacteria, causing deleterious mutation effects or death of cells harboring the genome. In short, this study provides reference for future relevant studies and suggestions for improvement of the strategies employed. Bachelor of Science in Biological Sciences 2014-05-08T08:59:46Z 2014-05-08T08:59:46Z 2014 2014 Final Year Project (FYP) http://hdl.handle.net/10356/59590 en Nanyang Technological University 35 p. application/pdf |
| spellingShingle | DRNTU::Science::Biological sciences Chia, Pei Shan Replication of norovirus in cultured cells |
| title | Replication of norovirus in cultured cells |
| title_full | Replication of norovirus in cultured cells |
| title_fullStr | Replication of norovirus in cultured cells |
| title_full_unstemmed | Replication of norovirus in cultured cells |
| title_short | Replication of norovirus in cultured cells |
| title_sort | replication of norovirus in cultured cells |
| topic | DRNTU::Science::Biological sciences |
| url | http://hdl.handle.net/10356/59590 |
| work_keys_str_mv | AT chiapeishan replicationofnorovirusinculturedcells |