PiggyBac Transposon-Mediated Transgenesis in the Pacific Oyster (Crassostrea gigas) – First Time in Mollusks
As an effective method of transgenesis, the plasmid of PiggyBac transposon containing GFP (PiggyBac) transposon system has been widely used in various organisms but not yet in mollusks. In this work, piggyBac containing green fluorescent protein (GFP) was transferred into the Pacific oyster (Crassos...
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ftdoajarticles:oai:doaj.org/article:e8322376b32c4580b0f2865fbf8c5b4a 2023-05-15T15:58:01+02:00 PiggyBac Transposon-Mediated Transgenesis in the Pacific Oyster (Crassostrea gigas) – First Time in Mollusks Jun Chen Changlu Wu Baolu Zhang Zhongqiang Cai Lei Wei Zhuang Li Guangbin Li Ting Guo Yongchuan Li Wen Guo Xiaotong Wang 2018-07-01T00:00:00Z https://doi.org/10.3389/fphys.2018.00811 https://doaj.org/article/e8322376b32c4580b0f2865fbf8c5b4a EN eng Frontiers Media S.A. https://www.frontiersin.org/article/10.3389/fphys.2018.00811/full https://doaj.org/toc/1664-042X 1664-042X doi:10.3389/fphys.2018.00811 https://doaj.org/article/e8322376b32c4580b0f2865fbf8c5b4a Frontiers in Physiology, Vol 9 (2018) Pacific oyster transgenesis piggyBac transposons sperm-mediated gene transfer electroporation Physiology QP1-981 article 2018 ftdoajarticles https://doi.org/10.3389/fphys.2018.00811 2022-12-31T11:47:48Z As an effective method of transgenesis, the plasmid of PiggyBac transposon containing GFP (PiggyBac) transposon system has been widely used in various organisms but not yet in mollusks. In this work, piggyBac containing green fluorescent protein (GFP) was transferred into the Pacific oyster (Crassostrea gigas) by sperm-mediated gene transfer with or without electroporation. Fluorescent larvae were then observed and isolated under an inverted fluorescence microscope, and insertion of piggyBac was tested by polymerase chain reaction (PCR) using genomic DNA as template. Oyster larvae with green fluorescence were observed after transgenesis with or without electroporation, but electroporation increased the efficiency of sperm-mediated transgenesis. Subsequently, the recombinant piggyBac plasmid containing gGH (piggyBac-gGH) containing GFP and a growth hormone gene from orange-spotted grouper (gGH) was transferred into oysters using sperm mediation with electroporation, and fluorescent larvae were observed and isolated. The insertion of piggyBac-gGH was tested by PCR and genome walking analysis. PCR analysis indicated that piggyBac-gGH was transferred into oyster larvae; genome walking analysis further showed the detailed location where piggyBac-gGH was inserted in the oyster genome. This is the first time that piggyBac transposon-mediated transgenesis has been applied in mollusks. Article in Journal/Newspaper Crassostrea gigas Pacific oyster Directory of Open Access Journals: DOAJ Articles Pacific Frontiers in Physiology 9 |
institution |
Open Polar |
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Directory of Open Access Journals: DOAJ Articles |
op_collection_id |
ftdoajarticles |
language |
English |
topic |
Pacific oyster transgenesis piggyBac transposons sperm-mediated gene transfer electroporation Physiology QP1-981 |
spellingShingle |
Pacific oyster transgenesis piggyBac transposons sperm-mediated gene transfer electroporation Physiology QP1-981 Jun Chen Changlu Wu Baolu Zhang Zhongqiang Cai Lei Wei Zhuang Li Guangbin Li Ting Guo Yongchuan Li Wen Guo Xiaotong Wang PiggyBac Transposon-Mediated Transgenesis in the Pacific Oyster (Crassostrea gigas) – First Time in Mollusks |
topic_facet |
Pacific oyster transgenesis piggyBac transposons sperm-mediated gene transfer electroporation Physiology QP1-981 |
description |
As an effective method of transgenesis, the plasmid of PiggyBac transposon containing GFP (PiggyBac) transposon system has been widely used in various organisms but not yet in mollusks. In this work, piggyBac containing green fluorescent protein (GFP) was transferred into the Pacific oyster (Crassostrea gigas) by sperm-mediated gene transfer with or without electroporation. Fluorescent larvae were then observed and isolated under an inverted fluorescence microscope, and insertion of piggyBac was tested by polymerase chain reaction (PCR) using genomic DNA as template. Oyster larvae with green fluorescence were observed after transgenesis with or without electroporation, but electroporation increased the efficiency of sperm-mediated transgenesis. Subsequently, the recombinant piggyBac plasmid containing gGH (piggyBac-gGH) containing GFP and a growth hormone gene from orange-spotted grouper (gGH) was transferred into oysters using sperm mediation with electroporation, and fluorescent larvae were observed and isolated. The insertion of piggyBac-gGH was tested by PCR and genome walking analysis. PCR analysis indicated that piggyBac-gGH was transferred into oyster larvae; genome walking analysis further showed the detailed location where piggyBac-gGH was inserted in the oyster genome. This is the first time that piggyBac transposon-mediated transgenesis has been applied in mollusks. |
format |
Article in Journal/Newspaper |
author |
Jun Chen Changlu Wu Baolu Zhang Zhongqiang Cai Lei Wei Zhuang Li Guangbin Li Ting Guo Yongchuan Li Wen Guo Xiaotong Wang |
author_facet |
Jun Chen Changlu Wu Baolu Zhang Zhongqiang Cai Lei Wei Zhuang Li Guangbin Li Ting Guo Yongchuan Li Wen Guo Xiaotong Wang |
author_sort |
Jun Chen |
title |
PiggyBac Transposon-Mediated Transgenesis in the Pacific Oyster (Crassostrea gigas) – First Time in Mollusks |
title_short |
PiggyBac Transposon-Mediated Transgenesis in the Pacific Oyster (Crassostrea gigas) – First Time in Mollusks |
title_full |
PiggyBac Transposon-Mediated Transgenesis in the Pacific Oyster (Crassostrea gigas) – First Time in Mollusks |
title_fullStr |
PiggyBac Transposon-Mediated Transgenesis in the Pacific Oyster (Crassostrea gigas) – First Time in Mollusks |
title_full_unstemmed |
PiggyBac Transposon-Mediated Transgenesis in the Pacific Oyster (Crassostrea gigas) – First Time in Mollusks |
title_sort |
piggybac transposon-mediated transgenesis in the pacific oyster (crassostrea gigas) – first time in mollusks |
publisher |
Frontiers Media S.A. |
publishDate |
2018 |
url |
https://doi.org/10.3389/fphys.2018.00811 https://doaj.org/article/e8322376b32c4580b0f2865fbf8c5b4a |
geographic |
Pacific |
geographic_facet |
Pacific |
genre |
Crassostrea gigas Pacific oyster |
genre_facet |
Crassostrea gigas Pacific oyster |
op_source |
Frontiers in Physiology, Vol 9 (2018) |
op_relation |
https://www.frontiersin.org/article/10.3389/fphys.2018.00811/full https://doaj.org/toc/1664-042X 1664-042X doi:10.3389/fphys.2018.00811 https://doaj.org/article/e8322376b32c4580b0f2865fbf8c5b4a |
op_doi |
https://doi.org/10.3389/fphys.2018.00811 |
container_title |
Frontiers in Physiology |
container_volume |
9 |
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1766393732712628224 |