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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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Autores principales: Prasetyo, Frengky H.H., Sugiharto, Bambang, Ermawati, Netty
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Academy of Scientific Research and Technology, Egypt 2018
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.
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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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