title: Reduced protease activity in transformed rice cell suspension cultures expressing a proteinase inhibitor.
abstract: 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 alpha-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.
authors.affiliation: Division of Biological Sciences and the Research Center for Bioactive Materials, Chonbuk National University, Jeonju 561-756, Republic of Korea.
authors.forename: Tae-Geum
authors.initials: TG
authors.lastname: Kim
authors.forename: Hyang-Mi
authors.initials: HM
authors.lastname: Kim
authors.forename: Ho-Jin
authors.initials: HJ
authors.lastname: Lee
authors.forename: Yun-Ji
authors.initials: YJ
authors.lastname: Shin
authors.forename: Tae-Ho
authors.initials: TH
authors.lastname: Kwon
authors.forename: Nan-Ju
authors.initials: NJ
authors.lastname: Lee
authors.forename: Yong-Suk
authors.initials: YS
authors.lastname: Jang
authors.forename: Moon-Sik
authors.initials: MS
authors.lastname: Yang
mesh_terms: D001483:Base Sequence; D002478:Cells, Cultured; D017931:DNA Primers / Q000235:genetics; D018744:DNA, Plant / Q000235:genetics; D015870:Gene Expression; D017343:Genes, Plant; D005822:Genetic Vectors; D012275:Oryza / Q000235:genetics* / Q000378:metabolism*; D010447:Peptide Hydrolases / Q000378:metabolism*; D010940:Plant Proteins / Q000235:genetics* / Q000378:metabolism*; D030821:Plants, Genetically Modified; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D014026:Nicotiana / Q000235:genetics / Q000378:metabolism; D014170:Transformation, Genetic
chemical_list: D017931:DNA Primers; D018744:DNA, Plant; C075489:PI-I protein, Nicotiana tabacum; D010940:Plant Proteins; D011994:Recombinant Proteins; D010447:Peptide Hydrolases