Isolation and regeneration of transiently transformed protoplasts from gametophytic blades of the marine red alga Porphyra yezoensis
Despite the recent progress of transient gene expression systems in a red alga Porphyra yezoensis by particle bombardment, a stable transformation system has yet to establish in any marine red macrophytes. One of the reasons of the difficulty in genetic transformation in red algae is the lack of sys...
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Pontificia Universidad Católica de Valparaíso
2010
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oai:scielo:S0717-345820100002000082010-04-28Isolation and regeneration of transiently transformed protoplasts from gametophytic blades of the marine red alga Porphyra yezoensisTakahashi,MegumuUji,ToshikiSaga,NaotsuneMikami,Koji allantoin protoplast P. yezoensis regeneration transient gene expression Despite the recent progress of transient gene expression systems in a red alga Porphyra yezoensis by particle bombardment, a stable transformation system has yet to establish in any marine red macrophytes. One of the reasons of the difficulty in genetic transformation in red algae is the lack of systems to select and isolate transformed cells from gametophytic blades. Thus, toward the establishment of the stable transformation system in P. yezoensis, we have developed a procedure by which transiently transformed gametophytic cells were prepared from particle bombarded-gametophytic blade as regeneratable protoplasts. Using mixture of marine bacterial enzymes, yield of protoplasts was high as reported elsewhere; however, these protoplasts did not develop. In contrast, protoplasts prepared from gametophytes treated with allantoin were normally developed, in which the overexpression of a β-glucuronidase reporter gene had no effect on the regeneration of protoplasts. Therefore, the use of allantoin in protoplast preparation sheds a new light on the realization of an efficient isolation and selection of study transformed cells from gametophytic blades.info:eu-repo/semantics/openAccessPontificia Universidad Católica de ValparaísoElectronic Journal of Biotechnology v.13 n.2 20102010-03-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582010000200008en10.4067/S0717-34582010000200008 |
| institution |
Scielo Chile |
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Scielo Chile |
| language |
English |
| topic |
allantoin protoplast P. yezoensis regeneration transient gene expression |
| spellingShingle |
allantoin protoplast P. yezoensis regeneration transient gene expression Takahashi,Megumu Uji,Toshiki Saga,Naotsune Mikami,Koji Isolation and regeneration of transiently transformed protoplasts from gametophytic blades of the marine red alga Porphyra yezoensis |
| description |
Despite the recent progress of transient gene expression systems in a red alga Porphyra yezoensis by particle bombardment, a stable transformation system has yet to establish in any marine red macrophytes. One of the reasons of the difficulty in genetic transformation in red algae is the lack of systems to select and isolate transformed cells from gametophytic blades. Thus, toward the establishment of the stable transformation system in P. yezoensis, we have developed a procedure by which transiently transformed gametophytic cells were prepared from particle bombarded-gametophytic blade as regeneratable protoplasts. Using mixture of marine bacterial enzymes, yield of protoplasts was high as reported elsewhere; however, these protoplasts did not develop. In contrast, protoplasts prepared from gametophytes treated with allantoin were normally developed, in which the overexpression of a β-glucuronidase reporter gene had no effect on the regeneration of protoplasts. Therefore, the use of allantoin in protoplast preparation sheds a new light on the realization of an efficient isolation and selection of study transformed cells from gametophytic blades. |
| author |
Takahashi,Megumu Uji,Toshiki Saga,Naotsune Mikami,Koji |
| author_facet |
Takahashi,Megumu Uji,Toshiki Saga,Naotsune Mikami,Koji |
| author_sort |
Takahashi,Megumu |
| title |
Isolation and regeneration of transiently transformed protoplasts from gametophytic blades of the marine red alga Porphyra yezoensis |
| title_short |
Isolation and regeneration of transiently transformed protoplasts from gametophytic blades of the marine red alga Porphyra yezoensis |
| title_full |
Isolation and regeneration of transiently transformed protoplasts from gametophytic blades of the marine red alga Porphyra yezoensis |
| title_fullStr |
Isolation and regeneration of transiently transformed protoplasts from gametophytic blades of the marine red alga Porphyra yezoensis |
| title_full_unstemmed |
Isolation and regeneration of transiently transformed protoplasts from gametophytic blades of the marine red alga Porphyra yezoensis |
| title_sort |
isolation and regeneration of transiently transformed protoplasts from gametophytic blades of the marine red alga porphyra yezoensis |
| publisher |
Pontificia Universidad Católica de Valparaíso |
| publishDate |
2010 |
| url |
http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582010000200008 |
| work_keys_str_mv |
AT takahashimegumu isolationandregenerationoftransientlytransformedprotoplastsfromgametophyticbladesofthemarineredalgaporphyrayezoensis AT ujitoshiki isolationandregenerationoftransientlytransformedprotoplastsfromgametophyticbladesofthemarineredalgaporphyrayezoensis AT saganaotsune isolationandregenerationoftransientlytransformedprotoplastsfromgametophyticbladesofthemarineredalgaporphyrayezoensis AT mikamikoji isolationandregenerationoftransientlytransformedprotoplastsfromgametophyticbladesofthemarineredalgaporphyrayezoensis |
| _version_ |
1718441813287632896 |