Gene Transfer through Electroporation of Sweet Potato Intact Cells

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Title:	Gene Transfer through Electroporation of Sweet Potato Intact Cells 甘藷完整細胞之電擊穿孔基因轉移
Authors:	林育瑰黃士穎
Contributors:	農學院
Keywords:	電擊穿孔單細胞甘藷基因暫時表現 Electroporation Cells Sweet potato Transient gene expression
Date:	1998-10
Issue Date:	2016-10-28 15:03:54 (UTC+8)
Abstract:	本研究報導公電擊穿孔的方涇，將含有gus報導基因之pBI221質體DNA導入甘藷細胞中。實驗結果發現當先以1%(v/v)巰基乙醇(β-mercaptoethanol)處理甘藷細胞，再以500伏特電壓及490微法拉第之電容，於脈衝寬度500毫秒等之電擊條件下電擊兩次，可發現報導基因於被電擊之細胞內有暫時表現之結果。因此藉由電擊穿孔以轉移基因之方法，不但可以應用於原生質體也可應用於具完整細胞壁的單細胞。 Cells isolated from sweet potato leaves were electroporated to examine transient expression of gus gene. Plasmid pBI221 DAN containing gus reporter gene was used to examine whether electroporated cells were transformed. Pretreatment with β-Mercaptoethanol (1%, v/v) and electroporated with two pulses at 500 V cm-1 and 490μF capacitance with a pulse length of 500 ms reduced cell viability to 54% and there were detectable β-glucuronidase activity in the electroporated cells. No β-glucuronidase activity was found for electroporated sweet potato cells without the pretreatment with β-Mercaptoethanol. The highest β-glucuronidase activity in the electroporated cells was observed when cells were pretreated with 1.5% (v/v) β-Mercaptoethanol. The levels of transient expression of gus gene was improved by an increase in plasmid concentration. This study demonstrated that sweet potato cells with intact cell wall rather than protoplast without cell wall can be electroporated and reporter gene expressed. This method of intact cell electroporation should be applicable to the plant species that protoplast isolation is difficult.
Relation:	華岡農科學報 8 民87.10 頁89-97
Appears in Collections:	[College of Agriculture] Hwa Kang journal of Agriculture

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