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Metabolic regulation of α-amylase gene expression in transgenic cell cultures of rice (Oryza sativa L.) (1993)

Huang, Ning ; Chandler, John ; Thomas, Bruce R. ; [et al.]

Springer

Plant molecular biology 23 (1993), S. 737-747

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ISSN: 1573-5028

Keywords: α-amylase ; transgenic cell culture ; gene expression ; metabolic regulation ; Oryza sativa ; transcription

Source: Springer Online Journal Archives 1860-2000

Topics: Biology

Notes: Abstract Expression of two genes in the α-amylase gene family is controlled by metabolic regulation in rice cultured cells. The levels of RAmy3D and RAmy3E mRNAs in rice cultured cells are inversely related to the concentration of sugar in the culture medium. Other genes in the rice α-amylase gene family have little or no expression in cultured cells; these expression levels are not controlled by metabolic regulation. A RAmy3D promoter/GUS gene fusion was metabolically regulated in the transgenic rice cell line 3DG, just as the endogenous RAmy3D gene is regulated. An assay of GUS enzyme activity in 3DG cells demonstrated that RAmy3D/GUS expression is repressed when sugar is present in the culture medium and induced when sugar is removed from the medium. The 942 bp fragment of the RAmy3D promoter that was linked to the coding region of the GUS reporter gene thus contains all of the regulatory sequences necessary for metabolic regulation of the gene.

Type of Medium: Electronic Resource

URL: http://dx.doi.org/10.1007/BF00021529

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Synthesis and secretion of α-amylase by rice callus: Evidence for differential gene expression (1991)

Simmons, Carl R. ; Huang, Ning ; Cao, Yicheng ; [et al.]

New York, NY [u.a.] : Wiley-Blackwell

Biotechnology and Bioengineering 38 (1991), S. 545-551

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ISSN: 0006-3592

Keywords: rice ; α-amylase ; callus ; mRNA ; and 2,4-dichlorophenoxyacetic acid (2,4-D) ; plant cell fermentation ; Chemistry ; Biochemistry and Biotechnology

Source: Wiley InterScience Backfile Collection 1832-2000

Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology

Notes: Rice seed callus expressed and secreted α-amylase at high levels. Twenty percent of the protein secreted by the callus was αamylase. The callus secreted about 840 μg α-amylase with 10.9 × 103 units of activity per gram dry weight callus per day. The α-amylase from callus exhibited a more complex isoform pattern than the germinating seed α-amylase. In addition, the level of mRNA expression by the five α-amylase gene groups was markedly different between callus and the germinating seed. The rice callus culture has features which it attractive as a potential system for expression proteins in plant cell fermentation systems.

Additional Material: 3 Ill.