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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ftpubmed:oai:pubmedcentral.nih.gov:6054966 2023-05-15T15:57:59+02:00 PiggyBac Transposon-Mediated Transgenesis in the Pacific Oyster (Crassostrea gigas) – First Time in Mollusks Chen, Jun Wu, Changlu Zhang, Baolu Cai, Zhongqiang Wei, Lei Li, Zhuang Li, Guangbin Guo, Ting Li, Yongchuan Guo, Wen Wang, Xiaotong 2018-07-16 http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6054966/ http://www.ncbi.nlm.nih.gov/pubmed/30061837 https://doi.org/10.3389/fphys.2018.00811 en eng Frontiers Media S.A. http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6054966/ http://www.ncbi.nlm.nih.gov/pubmed/30061837 http://dx.doi.org/10.3389/fphys.2018.00811 Copyright © 2018 Chen, Wu, Zhang, Cai, Wei, Li, Li, Guo, Li, Guo and Wang. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. CC-BY Physiology Text 2018 ftpubmed https://doi.org/10.3389/fphys.2018.00811 2018-08-05T00:27:24Z 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. Text Crassostrea gigas Pacific oyster PubMed Central (PMC) Pacific Frontiers in Physiology 9 |
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topic |
Physiology |
spellingShingle |
Physiology Chen, Jun Wu, Changlu Zhang, Baolu Cai, Zhongqiang Wei, Lei Li, Zhuang Li, Guangbin Guo, Ting Li, Yongchuan Guo, Wen Wang, Xiaotong PiggyBac Transposon-Mediated Transgenesis in the Pacific Oyster (Crassostrea gigas) – First Time in Mollusks |
topic_facet |
Physiology |
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 |
Text |
author |
Chen, Jun Wu, Changlu Zhang, Baolu Cai, Zhongqiang Wei, Lei Li, Zhuang Li, Guangbin Guo, Ting Li, Yongchuan Guo, Wen Wang, Xiaotong |
author_facet |
Chen, Jun Wu, Changlu Zhang, Baolu Cai, Zhongqiang Wei, Lei Li, Zhuang Li, Guangbin Guo, Ting Li, Yongchuan Guo, Wen Wang, Xiaotong |
author_sort |
Chen, Jun |
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 |
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6054966/ http://www.ncbi.nlm.nih.gov/pubmed/30061837 https://doi.org/10.3389/fphys.2018.00811 |
geographic |
Pacific |
geographic_facet |
Pacific |
genre |
Crassostrea gigas Pacific oyster |
genre_facet |
Crassostrea gigas Pacific oyster |
op_relation |
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6054966/ http://www.ncbi.nlm.nih.gov/pubmed/30061837 http://dx.doi.org/10.3389/fphys.2018.00811 |
op_rights |
Copyright © 2018 Chen, Wu, Zhang, Cai, Wei, Li, Li, Guo, Li, Guo and Wang. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
op_rightsnorm |
CC-BY |
op_doi |
https://doi.org/10.3389/fphys.2018.00811 |
container_title |
Frontiers in Physiology |
container_volume |
9 |
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1766393704315092992 |