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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
刊名	WORLD JOURNAL OF MICROBIOLOGY & BIOTECHNOLOGY
	2010-09-01
卷号	26期号:9页码:1653-1657
关键词	Glass-bead Method Green Fluorescent Protein Platymonas Subcordiformis Protoplast Transgenosis
ISSN号	0959-3993
DOI	10.1007/s11274-010-0342-6
文献子类	Article
英文摘要	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.
学科主题	Biotechnology & Applied Microbiology
URL标识	查看原文
语种	英语
WOS记录号	WOS:000280642800013
公开日期	2010-12-24
内容类型	期刊论文
源URL	[http://ir.qdio.ac.cn/handle/337002/5949]
专题	海洋研究所_实验海洋生物学重点实验室 海洋研究所_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
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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