Institutional Repository of Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences
| Transformation of Platymonas (Tetraselmis) subcordiformis (Prasinophyceae, Chlorophyta) by agitation with glass beads | |
Cui, Yulin1,2; Wang, Jinfeng1,2 ; Jiang, Peng1; Bian, Shuguang3; Qin, Song1
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| 2010-09-01 | |
| 发表期刊 | WORLD JOURNAL OF MICROBIOLOGY & BIOTECHNOLOGY
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| ISSN | 0959-3993 |
| 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 |
| DOI | 10.1007/s11274-010-0342-6 |
| URL | 查看原文 |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS记录号 | WOS:000280642800013 |
| 引用统计 | |
| 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 |
| 推荐引用方式 GB/T 7714 | 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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