Reduced protease activity in transformed rice cell suspension cultures expressing a proteinase inhibitor

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• 作者：Tae-Geum Kim ; Hyang-Mi Kim ; Ho-Jin Lee ; Yun-Ji Shin ; Tae-Ho Kwon ; Nan-Ju Lee ; Yong-Suk Jang ; Moon-Sik Yang
• 关键词：Cell suspension culture ; Rice ; Serine proteinase inhibitor II
• 刊名：Protein Expression and Purification
• 出版年：2007
• 出版时间：June 2007
• 年：2007
• 卷：53
• 期：2
• 页码：270-274
• 全文大小：479 K

文摘

In this study, we synthesized a synthetic serine proteinase inhibitor II gene (sPI-II) that harbored the chymotrypsin and trypsin inhibitor domains of the PI-II gene from Nicotiana alata. In an effort to reduce protease activity in a rice cell suspension culture, we first synthesized sPI-II using overlap PCR and then introduced the gene into a rice calli (Oryza sativa L. cv. Dongin) by particle bombardment-mediated transformation. The sPI-II gene was under the control of a rice α-amylase 3D promoter induced by sugar starvation. To verify the integration and expression of the sPI-II gene in the transformed rice cells, we employed genomic DNA PCR amplification and Northern blot analysis, respectively. The relative protease activity of the transformed cell suspension culture was reduced to approximately 23 % when compared to the non-transformed culture. This indicates that a transformed suspension culture system expressing a proteinase inhibitor, may be a useful tool to protect against recombinant protein losses resulting from extracellular proteases.