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Cloning, transformation and expression of cell cycle-associated protein kinase OsWee1 in indica rice (Oryza sativa L.)
The development process of seed in plants is a cycle of cells which occur gradually and regularly. One of the genes involved in controling this stage is the Wee1 gene. Wee1 encode protein kinase which plays an important role in phosphorylation, inactivation of cyclin-dependent kinase 1 (CDK1)-cyclin...
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Academy of Scientific Research and Technology, Egypt
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6353929/ https://www.ncbi.nlm.nih.gov/pubmed/30733775 http://dx.doi.org/10.1016/j.jgeb.2018.10.003 |
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author | Prasetyo, Frengky H.H. Sugiharto, Bambang Ermawati, Netty |
author_facet | Prasetyo, Frengky H.H. Sugiharto, Bambang Ermawati, Netty |
author_sort | Prasetyo, Frengky H.H. |
collection | PubMed |
description | The development process of seed in plants is a cycle of cells which occur gradually and regularly. One of the genes involved in controling this stage is the Wee1 gene. Wee1 encode protein kinase which plays an important role in phosphorylation, inactivation of cyclin-dependent kinase 1 (CDK1)-cyclin (CYC) and inhibiting cell division at mitotic phase. The Overexpression of Wee1 leads to delaying entry into mitotic phase, resulting in enlargement of cell size due to suppression of cell division. Accordingly, the cloning and overexpressing of Wee1 in rice plant is important aim of this research in achieving better quantity and quality of future rice. The main objective of this present study is to cloning and generate transgenic rice plants overexpressing of Wee1 gene. Wee1 was isolated from cDNA of indica rice (Oryza sativa), called OsWee1. The full length of OsWee1 was 1239 bp in size and successfully inserted into plant expression vector pRI101ON. Seven-day-old rice seedlings were prepared for transformation of OsWee1 gene using Agrobacterium-mediated transformation method. Four positive transgenic lines were identified through the presence of kanamycin resistance gene (nptII) using genomic PCR analysis. Southern blot analysis result provides evidence that four independent rice transformants contained one to three rearranged transgene copies. Further screening in transgenic rice generation is needed in order to obtain stable expression of OsWee1. |
format | Online Article Text |
id | pubmed-6353929 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Academy of Scientific Research and Technology, Egypt |
record_format | MEDLINE/PubMed |
spelling | pubmed-63539292019-02-07 Cloning, transformation and expression of cell cycle-associated protein kinase OsWee1 in indica rice (Oryza sativa L.) Prasetyo, Frengky H.H. Sugiharto, Bambang Ermawati, Netty J Genet Eng Biotechnol Plant Biotechnology The development process of seed in plants is a cycle of cells which occur gradually and regularly. One of the genes involved in controling this stage is the Wee1 gene. Wee1 encode protein kinase which plays an important role in phosphorylation, inactivation of cyclin-dependent kinase 1 (CDK1)-cyclin (CYC) and inhibiting cell division at mitotic phase. The Overexpression of Wee1 leads to delaying entry into mitotic phase, resulting in enlargement of cell size due to suppression of cell division. Accordingly, the cloning and overexpressing of Wee1 in rice plant is important aim of this research in achieving better quantity and quality of future rice. The main objective of this present study is to cloning and generate transgenic rice plants overexpressing of Wee1 gene. Wee1 was isolated from cDNA of indica rice (Oryza sativa), called OsWee1. The full length of OsWee1 was 1239 bp in size and successfully inserted into plant expression vector pRI101ON. Seven-day-old rice seedlings were prepared for transformation of OsWee1 gene using Agrobacterium-mediated transformation method. Four positive transgenic lines were identified through the presence of kanamycin resistance gene (nptII) using genomic PCR analysis. Southern blot analysis result provides evidence that four independent rice transformants contained one to three rearranged transgene copies. Further screening in transgenic rice generation is needed in order to obtain stable expression of OsWee1. Academy of Scientific Research and Technology, Egypt 2018-12 2018-12-07 /pmc/articles/PMC6353929/ /pubmed/30733775 http://dx.doi.org/10.1016/j.jgeb.2018.10.003 Text en © 2018 Production and hosting by Elsevier B.V. on behalf of Academy of Scientific Research & Technology. http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Plant Biotechnology Prasetyo, Frengky H.H. Sugiharto, Bambang Ermawati, Netty Cloning, transformation and expression of cell cycle-associated protein kinase OsWee1 in indica rice (Oryza sativa L.) |
title | Cloning, transformation and expression of cell cycle-associated protein kinase OsWee1 in indica rice (Oryza sativa L.) |
title_full | Cloning, transformation and expression of cell cycle-associated protein kinase OsWee1 in indica rice (Oryza sativa L.) |
title_fullStr | Cloning, transformation and expression of cell cycle-associated protein kinase OsWee1 in indica rice (Oryza sativa L.) |
title_full_unstemmed | Cloning, transformation and expression of cell cycle-associated protein kinase OsWee1 in indica rice (Oryza sativa L.) |
title_short | Cloning, transformation and expression of cell cycle-associated protein kinase OsWee1 in indica rice (Oryza sativa L.) |
title_sort | cloning, transformation and expression of cell cycle-associated protein kinase oswee1 in indica rice (oryza sativa l.) |
topic | Plant Biotechnology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6353929/ https://www.ncbi.nlm.nih.gov/pubmed/30733775 http://dx.doi.org/10.1016/j.jgeb.2018.10.003 |
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