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Transformation of Platymonas (Tetraselmis) subcordiformis (Prasinophyceae, Chlorophyta) by agitation with glass beads
Cui, Yulin1,2; Wang, Jinfeng1,2; Jiang, Peng1; Bian, Shuguang3; Qin, Song1
2010-09-01
发表期刊WORLD JOURNAL OF MICROBIOLOGY & BIOTECHNOLOGY
ISSN0959-3993
卷号26期号:9页码:1653-1657
文章类型Article
摘要A transient transformation system for the unicellular marine green alga, Platymonas subcordiformis, was established in this study. We introduced the pEGFP-N1 vector into P. subcordiformis with a glass bead method. P. subcordiformis was incubated in cell wall lytic enzymes (abalone acetone powder and cellulase solutions) to degrade the cell wall. The applicable conditions for production of viable protoplasts were pH 6.5, 25 degrees C, and 3 h of enzyme treatment. The protoplast yield was 61.2% when P. subcordiformis cells were added to the enzyme solution at a concentration of 10(7) cell ml(-1). The protoplasts were immediately transformed with the pEGFP-N1 vector using glass-bead method. The transformation frequency was about 10(-5), and there was no GFP activity observed in either the negative or the blank controls. This study indicated that GFP was a sensitively transgenic reporter for P. subcordiformis, and the method of cell wall enzymolysis followed by glass bead agitation was applicable for the transformation of P. subcordiformis.; A transient transformation system for the unicellular marine green alga, Platymonas subcordiformis, was established in this study. We introduced the pEGFP-N1 vector into P. subcordiformis with a glass bead method. P. subcordiformis was incubated in cell wall lytic enzymes (abalone acetone powder and cellulase solutions) to degrade the cell wall. The applicable conditions for production of viable protoplasts were pH 6.5, 25 degrees C, and 3 h of enzyme treatment. The protoplast yield was 61.2% when P. subcordiformis cells were added to the enzyme solution at a concentration of 10(7) cell ml(-1). The protoplasts were immediately transformed with the pEGFP-N1 vector using glass-bead method. The transformation frequency was about 10(-5), and there was no GFP activity observed in either the negative or the blank controls. This study indicated that GFP was a sensitively transgenic reporter for P. subcordiformis, and the method of cell wall enzymolysis followed by glass bead agitation was applicable for the transformation of P. subcordiformis.
关键词Glass-bead Method Green Fluorescent Protein Platymonas Subcordiformis Protoplast Transgenosis
学科领域Biotechnology & Applied Microbiology
DOI10.1007/s11274-010-0342-6
URL查看原文
收录类别SCI
语种英语
WOS记录号WOS:000280642800013
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被引频次:12[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://ir.qdio.ac.cn/handle/337002/5948
专题实验海洋生物学重点实验室
海洋生态与环境科学重点实验室
作者单位1.Chinese Acad Sci, Key Lab Expt Marine Biol, Inst Oceanol, Qingdao 266071, Peoples R China
2.Chinese Acad Sci, Grad Univ, Beijing 100049, Peoples R China
3.Nanjing Agr Univ, Key Lab Marine Biol Jiangsu Prov, Nanjing 210095, Peoples R China
第一作者单位中国科学院海洋研究所
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Cui, Yulin,Wang, Jinfeng,Jiang, Peng,et al. Transformation of Platymonas (Tetraselmis) subcordiformis (Prasinophyceae, Chlorophyta) by agitation with glass beads[J]. WORLD JOURNAL OF MICROBIOLOGY & BIOTECHNOLOGY,2010,26(9):1653-1657.
APA Cui, Yulin,Wang, Jinfeng,Jiang, Peng,Bian, Shuguang,&Qin, Song.(2010).Transformation of Platymonas (Tetraselmis) subcordiformis (Prasinophyceae, Chlorophyta) by agitation with glass beads.WORLD JOURNAL OF MICROBIOLOGY & BIOTECHNOLOGY,26(9),1653-1657.
MLA Cui, Yulin,et al."Transformation of Platymonas (Tetraselmis) subcordiformis (Prasinophyceae, Chlorophyta) by agitation with glass beads".WORLD JOURNAL OF MICROBIOLOGY & BIOTECHNOLOGY 26.9(2010):1653-1657.
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