Isolation of an Efficient Actin Promoter for Use in Rice Transformation

McElroy, David; Zhang, Wanggen; Cao, Jian; Wü, Ray

doi:10.2307/3868928

Cited by 192 publications

(239 citation statements)

References 10 publications

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“…To confirm the enhancer function, SRS was inserted in one, two, and three tandem copies in the EcoRI site (Ϫ459 bp upstream of the transcription start site) of the rice Act1 promoter (52). The wild type and the SRS-containing Act1 promoters were fused upstream of Luc gene as shown in Fig.…”

Section: Srs Acts As a Transcriptional Enhancer In A Sugar-insensitivmentioning

confidence: 99%

Sugar Response Sequence in the Promoter of a Rice α-Amylase Gene Serves as a Transcriptional Enhancer

Lu¹,

Lim²,

Yu³

1998

Journal of Biological Chemistry

160

163

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Expression of ␣-amylase genes in both rice suspension cells and germinating embryos is repressed by sugars and the mechanism involves transcriptional regulation. The promoter of a rice ␣-amylase gene ␣Amy3 was analyzed by both loss-and gain-of-function studies and the major sugar response sequence (SRS) was located between 186 and 82 base pairs upstream of the transcription start site. The SRS conferred sugar responsiveness to a minimal promoter in an orientation-independent manner. It also converted a sugar-insensitive rice actin gene promoter into a sugar-sensitive promoter in a dosedependent manner. Linker-scan mutation studies identified three essential motifs: the GC box, the G box, and the TATCCA element, within the SRS. Sequences containing either the GC box plus G box or the TATCCA element each mediated sugar response, however, they acted synergistically to give a high level glucose starvation-induced expression. Nuclear proteins from rice suspension cells binding to the TATCCA element in a sequence-specific and sugar-dependent manner were identified. The TATCCA element is also an important component of the gibberellin response complex of the ␣-amylase genes in germinating cereal grains, suggesting that the regulation of ␣-amylase gene expression by sugar and hormone signals may share common regulatory machinery.Sugar repression of gene expression is a fundamental and ubiquitous regulatory system for adjusting to changes in nutrient availability in both prokaryotic and eukaryotic cells. In microorganisms, glucose or other rapidly metabolizable carbon sources repress the expression of genes that code for enzymes related to the metabolism of other carbon sources. Our understanding of the mechanisms of sugar repression has been based largely on studies of microorganisms. In the case of Escherichia coli, a model to explain at the molecular level, the mechanism of sugar repression has been determined (1, 2). The mechanism of glucose repression in yeast is more complicated and is less understood than it is in E. coli (3-5). Studies using Saccharomyces cerevisiae mutants have revealed many of the components involved in the response to carbon catabolite repression (5), but it is still unclear how all of these components interact to regulate transcription. A universal signaling pathway which leads to the regulation of all glucose-repressible genes has yet to be determined.As in microorganisms, sugar repression of gene expression also allows plant cells to cope effectively with changes in the carbon sources present in their environment. However, in multicellular plants, feedback repression by excess sugars provides an additional mechanism for maintaining an economical balance between supply (source) and demand (sink) for carbohydrate allocation and utilization among tissues and organs (6 -8). Despite the fact that sugar repression of gene expression is likely a central control mechanism mediating energy homeostasis and carbohydrate distribution in plants, the molecular mechanism of sugar feedback regulation remain...

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Section: Srs Acts As a Transcriptional Enhancer In A Sugar-insensitivmentioning

confidence: 99%

Sugar Response Sequence in the Promoter of a Rice α-Amylase Gene Serves as a Transcriptional Enhancer

Lu¹,

Lim²,

Yu³

1998

Journal of Biological Chemistry

160

163

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“…A high level of expression in transgenic grains becomes an important issue when the commercial production, application, or purification requires a high concentration of recombinant protein in the grain. Successful increases of recombinant protein synthesis and storage have been achieved by codon optimization of the transgene to a C ϩ G content of more than 60% (1), by selection of a stronger promoter (7), and by using a signal peptide to target the protein into the endoplasmic reticulum for deposition into protein bodies (1,2). A further or alternative way of increasing the amount of recombinant protein might be the transgenic expression of transcription factors that have the ability to transactivate the target genes by binding to nucleotide sequence motifs in the gene promoter (8)(9)(10)(11).…”

mentioning

confidence: 99%

Expression of the REB transcriptional activator in rice grains improves the yield of recombinant proteins whose genes are controlled by a Reb -responsive promoter

Yang¹,

Wu²,

Hwang³

et al. 2001

Proc. Natl. Acad. Sci. U.S.A.

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The gene encoding the rice transcription factor, REB (rice endosperm bZIP) was cloned from a bacterial artificial chromosome library of rice. The cloned 6,227-bp-long Reb gene is composed of six exons and five introns and is flanked by a 1.2-kb 5 promoter and a 1.2-kb 3 terminator region. The function of the Reb gene was explored by a transient assay by using a rice immature endosperm system. The effector constructs containing the native gene or fusion genes linking Reb to the rice actin (Act) or globulin (Glb) gene promoters and the reporter gene construct Glb-␤-glucuronidase (GUS) were used in this study. When these effector constructs were cotransferred with the reporter uidA gene encoding GUS under the control of the Glb promoter into immature rice endosperm cells, the Glb promoter was activated. The transient GUS expression was 2.0 to 2.5-fold higher with the effector construct than without. When the upstream activation sequence containing the GCCACGT(A͞C)AG motifs of the Glb promoter was deleted, the activation by REB was abolished. On the other hand, a gain-offunction experiment showed that inserting the upstream activation sequence into the glutelin-1 (Gt1) promoter made it responsive to activation by REB. When cotransformed with Reb gene, mature transgenic rice grains containing the human lysozyme gene driven by the Glb promoter produced 3.7-fold more lysozyme. Accumulation of recombinant lysozyme in mature seed ranged from 30.57 to 279.61 g⅐mg ؊1 total soluble protein in individual transformants from 30 independent transformation events. Thus, our results show that REB is not only a transcriptional activator, it can also be used to increase the expression of recombinant protein in transgenic rice grains.Reb gene ͉ lysozyme expression ͉ transgenic plant ͉ gain-of-function ͉ lossof-function

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“…The plate was placed 8 cm beneath the stopping plate of the gun with a layer of metal net (120-180 mesh per linear inch) 1.5 cm above the target materials for even dispersion of tungsten particles. Plasmids used in the experiments included: pActlD which contains the rice actin1 gene promoter linked to the GUS gene (McElroy et al 1990; a gift from Dr. R. Wu); pNG3 which contains lhe NOS promoter driving HPH gene (a gift from Dr. M. C. Van Montagu); and pMON410 which contains CaMV 35s promoter driving the HPH' gcne (Rogers et al 1987; a gift from Monsanto Company). All plasmids were purified by CsCl gradient centrifugation.…”

Section: Particle Bombarhentmentioning

confidence: 99%

An improved rice transformation system using the biolistic method

et al. 1993

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Abstract. Immature embryos and embryogenic calli of rice, both japonica and indica subspecies, were bombarded with tungsten particles coated with plasmid DNA that contained a gene encoding hygromycin phosphotransferase (HPH, conferring hygromycin resistance) driven by the CaMV 35s promoter or Agrobacterium tumefacieiis NOS promoter. Putatively transformed cell clusters were identified from the bombarded tissues 2 weeks after selection on hygromycin B. By separating these cell clusms from each other, and by stringent selection not only at the callus growth stage but also during regeneration and plantlet growth, the overall transformation and selection efficiencies were substantially improved over those previously reported. From the most responsive cultivar used in these studies, an average of one transgenic plant was produced from 1.3 immature embryos or from 5 pieces of embryogenic calli bombarded. Integration of the introduced gene into the plant genome, and inheritance to the offspring were demonstrated. By using this procedure, we have produced several hundred transgenic plants. The procedure described here provides a simple method for improving transformation and selection efficiencies in rice and may be applicable to other monocots.

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Isolation of an Efficient Actin Promoter for Use in Rice Transformation

Cited by 192 publications

References 10 publications

Sugar Response Sequence in the Promoter of a Rice α-Amylase Gene Serves as a Transcriptional Enhancer

Sugar Response Sequence in the Promoter of a Rice α-Amylase Gene Serves as a Transcriptional Enhancer

Expression of the REB transcriptional activator in rice grains improves the yield of recombinant proteins whose genes are controlled by a Reb -responsive promoter

An improved rice transformation system using the biolistic method

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