دورية أكاديمية
The Application of the Gibson Assembly Method in the Production of Two pKLS3 Vector-Derived Infectious Clones of Foot-and-Mouth Disease Virus
| العنوان: | The Application of the Gibson Assembly Method in the Production of Two pKLS3 Vector-Derived Infectious Clones of Foot-and-Mouth Disease Virus |
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| المؤلفون: | Ploypailin Semkum, Nattarat Thangthamniyom, Penpitcha Chankeeree, Challika Keawborisuth, Sirin Theerawatanasirikul, Porntippa Lekcharoensuk |
| المصدر: | Vaccines, Vol 11, Iss 6, p 1111 (2023) |
| بيانات النشر: | MDPI AG, 2023. |
| سنة النشر: | 2023 |
| المجموعة: | LCC:Medicine |
| مصطلحات موضوعية: | Gibson assembly, foot-and-mouth disease virus, reverse genetics, vaccine, infectious clone, Medicine |
| الوصف: | The construction of a full-length infectious clone, essential for molecular virological study and vaccine development, is quite a challenge for viruses with long genomes or possessing complex nucleotide sequence structures. Herein, we have constructed infectious clones of foot-and-mouth disease virus (FMDV) types O and A by joining each viral coding region with our pKLS3 vector in a single isothermal reaction using Gibson Assembly (GA). pKLS3 is a 4.3-kb FMDV minigenome. To achieve optimal conditions for the DNA joining, each FMDV coding sequence was divided into two overlapping fragments of approximately 3.8 and 3.2 kb, respectively. Both DNA fragments contain the introduced linker sequences for assembly with the linearized pKLS3 vector. FMDV infectious clones were produced upon directly transfecting the GA reaction into baby hamster kidney-21 (BHK-21) cells. After passing in BHK-21 cells, both rescued FMDVs (rO189 and rNP05) demonstrated growth kinetics and antigenicity similar to their parental viruses. Thus far, this is the first report on GA-derived, full-length infectious FMDV cDNA clones. This simple DNA assembly method and the FMDV minigenome would facilitate the construction of FMDV infectious clones and enable genetic manipulation for FMDV research and custom-made FMDV vaccine production. |
| نوع الوثيقة: | article |
| وصف الملف: | electronic resource |
| اللغة: | English |
| تدمد: | 11061111 2076-393X |
| Relation: | https://www.mdpi.com/2076-393X/11/6/1111; https://doaj.org/toc/2076-393X |
| DOI: | 10.3390/vaccines11061111 |
| URL الوصول: | https://doaj.org/article/9604114c98e04614b23c604bb1ba9824 |
| رقم الأكسشن: | edsdoj.9604114c98e04614b23c604bb1ba9824 |
| قاعدة البيانات: | Directory of Open Access Journals |
Full Text Finder | تدمد: | 11061111 2076393X |
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| DOI: | 10.3390/vaccines11061111 |