Genomic integration and germline transmission of plasmid injected into crustacean Daphnia magna eggs.

The water flea, Daphnia, has been the subject of study in ecology, evolution, and environmental sciences for decades. Over the last few years, expressed sequence tags and a genome sequence have been determined. In addition, functional approaches of overexpression and gene silencing based on microinj...

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Autores principales:	Yasuhiko Kato, Tomoaki Matsuura, Hajime Watanabe
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2012
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/2a6c5516fb7a4087bb5939d240de3e62
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spelling	oai:doaj.org-article:2a6c5516fb7a4087bb5939d240de3e622021-11-18T07:05:09ZGenomic integration and germline transmission of plasmid injected into crustacean Daphnia magna eggs.1932-620310.1371/journal.pone.0045318https://doaj.org/article/2a6c5516fb7a4087bb5939d240de3e622012-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23028929/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203The water flea, Daphnia, has been the subject of study in ecology, evolution, and environmental sciences for decades. Over the last few years, expressed sequence tags and a genome sequence have been determined. In addition, functional approaches of overexpression and gene silencing based on microinjection of RNAs into eggs have been established. However, the transient nature of these approaches prevents us from analyzing gene functions in later stages of development. To overcome this limitation, transgenesis would become a key tool. Here we report establishment of a transgenic line using microinjection of plasmid into Daphnia magna eggs. The green fluorescent protein (GFP) gene fused with the D. magna histone H2B gene under the control of a promoter/enhancer region of the elongation factor 1α-1 (EF1α-1) gene, EF1α-1::H2B-GFP, was used as a reporter providing high resolution visualization of active chromatin. Transgenic lines were obtained from 0.67% of the total fertile adults that survived the injections. One of the transgenic animals, which exhibited fluorescence in the nuclei of cells during embryogenesis and oogenesis, had two copies of EF1α-1::H2B-GFP in a head-to-tail array. This is the first report of a transgenesis technique in Daphnia and, together with emerging genome sequences, will be useful for advancing knowledge of the molecular biology of Daphnia.Yasuhiko KatoTomoaki MatsuuraHajime WatanabePublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 7, Iss 9, p e45318 (2012)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Yasuhiko Kato Tomoaki Matsuura Hajime Watanabe Genomic integration and germline transmission of plasmid injected into crustacean Daphnia magna eggs.
description	The water flea, Daphnia, has been the subject of study in ecology, evolution, and environmental sciences for decades. Over the last few years, expressed sequence tags and a genome sequence have been determined. In addition, functional approaches of overexpression and gene silencing based on microinjection of RNAs into eggs have been established. However, the transient nature of these approaches prevents us from analyzing gene functions in later stages of development. To overcome this limitation, transgenesis would become a key tool. Here we report establishment of a transgenic line using microinjection of plasmid into Daphnia magna eggs. The green fluorescent protein (GFP) gene fused with the D. magna histone H2B gene under the control of a promoter/enhancer region of the elongation factor 1α-1 (EF1α-1) gene, EF1α-1::H2B-GFP, was used as a reporter providing high resolution visualization of active chromatin. Transgenic lines were obtained from 0.67% of the total fertile adults that survived the injections. One of the transgenic animals, which exhibited fluorescence in the nuclei of cells during embryogenesis and oogenesis, had two copies of EF1α-1::H2B-GFP in a head-to-tail array. This is the first report of a transgenesis technique in Daphnia and, together with emerging genome sequences, will be useful for advancing knowledge of the molecular biology of Daphnia.
format	article
author	Yasuhiko Kato Tomoaki Matsuura Hajime Watanabe
author_facet	Yasuhiko Kato Tomoaki Matsuura Hajime Watanabe
author_sort	Yasuhiko Kato
title	Genomic integration and germline transmission of plasmid injected into crustacean Daphnia magna eggs.
title_short	Genomic integration and germline transmission of plasmid injected into crustacean Daphnia magna eggs.
title_full	Genomic integration and germline transmission of plasmid injected into crustacean Daphnia magna eggs.
title_fullStr	Genomic integration and germline transmission of plasmid injected into crustacean Daphnia magna eggs.
title_full_unstemmed	Genomic integration and germline transmission of plasmid injected into crustacean Daphnia magna eggs.
title_sort	genomic integration and germline transmission of plasmid injected into crustacean daphnia magna eggs.
publisher	Public Library of Science (PLoS)
publishDate	2012
url	https://doaj.org/article/2a6c5516fb7a4087bb5939d240de3e62
work_keys_str_mv	AT yasuhikokato genomicintegrationandgermlinetransmissionofplasmidinjectedintocrustaceandaphniamagnaeggs AT tomoakimatsuura genomicintegrationandgermlinetransmissionofplasmidinjectedintocrustaceandaphniamagnaeggs AT hajimewatanabe genomicintegrationandgermlinetransmissionofplasmidinjectedintocrustaceandaphniamagnaeggs
_version_	1718423967951224832

Genomic integration and germline transmission of plasmid injected into crustacean Daphnia magna eggs.

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