共查询到20条相似文献,搜索用时 250 毫秒
1.
K. A. Torbert M. Gopalraj S. L. Medberry N. E. Olszewski D. A. Somers 《Plant cell reports》1998,17(4):284-287
The Commelina yellow mottle virus (CoYMV) infects the monocot weed Commelina diffusa. The objective of this study was to investigate the transgene expression conferred by the CoYMV promoter in a monocot species.
Friable, embryogenic oat (Avena sativa L.) tissue cultures were stably transformed with the CoYMV promoter fused to the coding region of E. coli β-glucuronidase (uidA, GUS). Developmental and tissue-specific expression of the CoYMV-GUS construct was investigated in regenerated plants and
their progeny. Histochemical GUS staining was primarily localized in the vascular tissues of shoots, leaves, floral bracts
and in roots. While ovaries stained intensely, no staining was detected in anthers or the endosperm in mature seed. The scutellum
of mature and germinating seed exhibited GUS activity.
Received: 16 April 1997 / Revision received: 23 July 1997 / Accepted: 2 August 1997 相似文献
2.
Guar seed gum, consisting primarily of a high molecular weight galactomannan, is the most cost effective natural thickener,
having broad applications in the food, cosmetics, paper, pharmaceutical and petroleum industries. The properties of the polymer
can potentially be enhanced by genetic modification. Development of suitable endosperm-specific promoters for use in guar
is desirable for metabolic engineering of the seed gum. A ~1.6 kb guar mannan synthase (MS) promoter region has been isolated. The MS promoter sequence was fused with the GUS reporter gene and overexpressed in the heterologous species alfalfa (Medicago sativa). The potential strength and specificity of the MS promoter was compared with those of the constitutive 35S promoter and
the seed specific β-phaseolin promoter. Quantitative GUS assays revealed that the MS promoter directs GUS expression specifically in endosperm in transgenic alfalfa. Thus, the guar MS promoter could prove generally useful for directing endosperm-specific expression of transgenes in legume species. 相似文献
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Saidi Y Finka A Chakhporanian M Zrÿd JP Schaefer DG Goloubinoff P 《Plant molecular biology》2005,59(5):697-711
The ability to express tightly controlled amounts of endogenous and recombinant proteins in plant cells is an essential tool
for research and biotechnology. Here, the inducibility of the soybean heat-shock Gmhsp17.3B promoter was addressed in the moss Physcomitrella patens, using β-glucuronidase (GUS) and an F-actin marker (GFP-talin) as reporter proteins. In stably transformed moss lines, Gmhsp17.3B-driven GUS expression was extremely low at 25 °C. In contrast, a short non-damaging heat-treatment at 38 °C rapidly induced
reporter expression over three orders of magnitude, enabling GUS accumulation and the labelling of F-actin cytoskeleton in
all cell types and tissues. Induction levels were tightly proportional to the temperature and duration of the heat treatment,
allowing fine-tuning of protein expression. Repeated heating/cooling cycles led to the massive GUS accumulation, up to 2.3%
of the total soluble proteins. The anti-inflammatory drug acetyl salicylic acid (ASA) and the membrane-fluidiser benzyl alcohol
(BA) also induced GUS expression at 25 °C, allowing the production of recombinant proteins without heat-treatment. The Gmhsp17.3B promoter thus provides a reliable versatile conditional promoter for the controlled expression of recombinant proteins in
the moss P. patens. 相似文献
4.
A promoter fusion (Sh35) combining upstream regulatory regions from the maize Sh1 promoter with a truncated 35S promoter, Δ9035 (–90 to +8) has been compared with the original Sh1 promoter for its capacity to promote expression of the β-glucuronidase (GUS) gene in stably transformed tomato plants. For both promoters, very faint GUS expression was detected
in the vegetative tissues, and no expression was detected in the fruit pericarp tissues. However, in the seed, Sh1 promoted low GUS expression but Sh35 directed 25-fold higher GUS expression. For both constructs, the profile of GUS expression
was similar to that of endogenous sucrose synthase activity, but maximal GUS activity was reached 15 days after the peak of
sucrose synthase activity.
Received: 20 October 1998 / Revision received: 1 December 1998 / Accepted: 14 December 1998 相似文献
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A. M. Ramadan Hala F. Eissa F. M. El-Domyati O. M. Saleh N. E. Ibrahim M. Salama M. M. Mahfouz A. Bahieldin 《Plant Cell, Tissue and Organ Culture》2011,107(3):373-381
The uidA gene, encoding for β-glucuronidase (GUS), is the most frequently used reporter gene in plants. As a reporter enzyme, GUS
can be assayed both qualitatively and quantitatively. In wheat, there are numerous reports of failure in detecting GUS enzyme
activity in tissues of transgenic plants, while other reports have suggested presence of β-glucuronidase inhibitor(s) in wheat
tissues. In the present study, we show that the β-glucuronidase enzyme activity is not only tissue-specific but also genotype-dependent.
Our data demonstrate that the glucuronic acid could be the candidate inhibitor for β-glucuronidase enzyme activity in wheat
leaves and roots. It should be noted that the assays to detect β-glucuronidase enzyme activity in wheat should be interpreted
carefully. Based on the data of our present study, we recommend studying the chemical pathways, the unintended effects and
the possible loss-of-function of any candidate transgene prior to transformation experiments. 相似文献
9.
P. Arokiaraj H. Yeet Yeang K. Fong Cheong S. Hamzah H. Jones S. Coomber B. V. Charlwood 《Plant cell reports》1998,17(8):621-625
Hevea brasiliensis anther calli were genetically transformed using Agrobacterium GV2260 (p35SGUSINT) that harboured the β-glucuronidase (gus) and neomycin phosphotransferase (nptII) genes. β-Glucuronidase protein (GUS) was expressed in the leaves of kanamycin-resistant plants that were regnerated, and the presence
of the gene was confirmed by Southern analysis. GUS was also observed to be expressed in the latex and more importantly in
the serum fraction. Transverse sections of the leaf petiole from a transformed plant revealed GUS expression to be especially
enhanced in the phloem and laticifers. GUS expression was subsequently detected in every one of 194 plants representing three
successive vegetative cycles propagated from the original transformant. Transgenic Hevea could thus facilitate the continual production of foreign proteins expressed in the latex.
Received: 14 February 1997 / Revision received: 16 August 1997 / Accepted: 20 July 1997 相似文献
10.
The harvesting and storing of alfalfa is a routine practice in the agricultural industry worldwide. To investigate gene expression
in harvested alfalfa, cDNA from non-harvested and harvested plants in the field was subjected to subtractive hybridization
to identify, in particular, those genes that are induced by the harvesting treatment. One cDNA clone, named hi11, was isolated and analysed. The full length cDNA of the hi11 gene was cloned by RACE amplification. The hi11 gene, which has high homology to a putative protein of unknown function in Arabidopsis, was induced in alfalfa following
harvesting, a 38°C heat shock and a wounding treatment. Northern blot analysis confirmed that the expression patterns of hi11 in alfalfa in response to harvesting, heat shock, and wounding. In addition, genomic walking was performed to isolate the
5′ flanking region of the hi11 gene. The promoter of the hi11 gene was fused to the GUS reporter gene and transferred to Medicago truncatula and tobacco. In all transgenic plants of M. truncatula and tobacco, GUS gene expression was observed in harvested tissue, especially in the transgenic tobacco plants, but not in
the non-harvested control tissue. 相似文献
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Lori A. Allison György B. Kiss Petra Bauer Maryse Poiret Michèle Pierre Arnould Savouré Eva Kondorosi Adam Kondorosi 《Plant molecular biology》1993,21(2):375-380
In a search for plant genes expressed during early symbiotic interactions between Medicago sativa and Rhizobium meliloti, we have isolated and characterized two alfalfa genes which have strong sequence similarity to members of the Enod12 gene family of Pisum sativum. The M. sativa genes, MsEnod12A and B, encode putative protein products of 8066 Da and 12849 Da, respectively, each with a signal sequence at the N-terminus followed by a repetitive proline-rich region. Based on their expression during the initial period of nodule development, MsEnod12A and B are alfalfa early nodulin genes. 相似文献
13.
Arabidopsis plants transformed by promoter of A. thaliana serine racemase fused with β-glucuronidase (GUS) reporter gene showed strong GUS staining in elongating and developing cells
such as tip regions of primary and lateral roots, developing leaves, and shoot meristems. RT-PCR and digital northern hybridization
showed that expression of the serine racemase gene was not induced by l- and d-serine, light irradiation, biotic and abiotic stresses. 相似文献
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In silico analysis showed that the differentially expressed type 3 oil palm metallothionein-like genes MT3-A and MT3-B share at least 11 common putative promoter regulatory elements. The identified motifs include W-boxes, TATCCA element, binding
element for cytokinin response regulators and pollen-specific elements. A high degree of conservation was observed in their
genomic organisation where the coding regions are divided at two identical positions in both genes by two AT-rich introns.
Promoter activity of the MT3-B gene was analysed using a transient assay by bombarding oil palm tissue slices with a β-glucuronidase (GUS) gene construct
and a stable reporter assay by analysing GUS expression in transformed Arabidopsis thaliana plants. Transient expression analysis revealed MT3-B promoter activity in oil palm root tissues but not in fruit mesocarp at 12 weeks after anthesis and spear leaves. The T3
homozygous transgenic Arabidopsis plants, harbouring the MT3-B promoter/GUS construct, showed reporter activity in cotyledons and mature leaves with lower expression levels in root tissues.
The expression levels in the roots of the T3 homozygous transgenic plants increased five- and 2.5-folds when treated with
80 μM of Zn2+ and Fe2+, respectively. Altogether, these results indicate that the MT3-A and MT3-B promoter activities may be regulated by a variety of abiotic factors and MT3-B promoter may potentially be manipulated for use in plant genetic engineering for induced synthesis of gene product. 相似文献
15.
K. B. McIntosh J. L. Hulm L. W. Young P. C. Bonham-Smith 《Plant Molecular Biology Reporter》2004,22(1):53-61
Stable transformation ofArabidopsis thaliana is a lengthy process that involves up to 3 mo of plant growth and seed selection. We have developed a rapid, 3-wk transient
assay system to test the functionality ofcis-regulatory regions controlling expression of a reporter gene in plants before undertaking stable transformation. Two-week-oldArabidopsis seedlings were vacuum-infiltrated withAgrobacterium tumefaciens cultures carrying various upstream regulatory regions controllinguidA (β-glucuronidase [GUS]) expression. Seedlings were fixed and stained for GUS activity 3–5 d following infiltration. Regulatory
regions tested in this system include the cauliflower mosaic virus (CaMV)35S promoter, the upstream regulatory region of ribosomal protein geneL23A-1, and a temperature-inducible regulatory region (HSP101B) also fromArabidopsis. The percentage of seedlings positive for GUS activity varied depending on the construct used, with the CaMV35S promoter producing the highest number of GUS-positive seedlings. Temperature induction treatments elicited increased GUS
expression in seedlings transformed with theHSP101B regulatory region. Regardless of construct, GUS expression levels were higher in seedlings collected 5 d followingAgrobacterium infiltration than those collected 3–4 d postinfiltration. 相似文献
16.
In our previous study, we identified a Rosa chinensis heat shock protein (HSP) gene, RcHSP17.8, which was induced by abiotic stresses, such as high temperature and osmotic stress. To analyze the expression of RcHSP17.8 and the function of cis-acting elements in the promoter region, a 1,910 bp fragment of the upstream sequence of the RcHSP17.8 translation initiation codon and five promoter deletion fragments were fused to a β-glucuronidase (GUS) report gene. These
plasmids were transferred to Arabidopsis thaliana via Agrobacterium. GUS staining was seen in all the organs, especially in the vascular tissues after heat treatment. In transgenic Arabidopsis, GUS expression driven by the full length promoter was significantly higher under heat shock, but no GUS activity was detected
under other abiotic stresses. Deletion analysis indicated that the region from −178 to −771 was essential for the promoter’s
response to high temperature. 相似文献
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It is desirable that the expression of transgenes in genetically modified crops is restricted to the tissues requiring the
encoded activity. To this end, we have studied the ability of the heterologous ribulose-1,5-bisphosphate carboxylase/oxygenase
(Rubisco) small-subunit (SSU) gene promoters, RBCS3CP (0.8 kbp) from tomato (hycopersion esculentum Mill.) and SRS1P (1.5 kbp) from soybean (Glycine max [h.] Mers.), to drive expression of the β-glucuronidase (gusA) marker gene in apple (Malus pumila Mill.). Transgenic lines of cultivar Greensleeves were produced by Agrobacterium-mediated transformation and the level of gusA expression in the vegetative tissues of young plants was compared with that produced using the cauliflower mosaic virus (CaMV)
35S promoter. These quantitative GUS data were assessed for their relationship to the copy number of transgene loci. The precise
location of GUS activity in leaves was identified histochemically. The heterologous SSU promoters were active primarily in
the green vegetative tissues of apple, although activity in the roots was noticeably higher with the RBCS3C promoter than with the SRS1 promoter. The mean GUS activity in leaf tissue of the SSU promoter transgenics was approximately half that of plants containing
the CaMV 35S promoter. Histochemical analysis demonstrated that GUS activity was localised to the mesophyll and palisade cells
of the leaf. The influence of light on expression was also determined. The activity of the SRS1 promoter was strictly dependent on light, whereas that of the RBCS3C promoter appeared not to be. Both SSU promoters would be suitable for the expression of transgenes in green photosynthetic
tissues of apple.
Received: 15 June 1999 / Accepted: 12 August 1999 相似文献
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Kazuko Yamaguchi-Shinozaki Kazuo Shinozaki 《Molecular & general genetics : MGG》1993,236(2-3):331-340
Summary We characterized the expression of genes that correspond to a cDNA clone, RD29, which is induced by desiccation, cold and high-salt conditions in Arabidopsis thaliana. Northern analysis of desiccation-induced expression revealed a two-step induction process. Early induction occurs within 20 min and secondary induction occurs 3 h after the start of desiccation. Exogenous abscisic acid (ABA) induces RD29 mRNA within 3 h. Two genes corresponding to RD29, rd29A and rd29B, are located in tandem in an 8 kb region of the Arabidopsis genome and encode hydrophilic proteins. Desiccation induces rd29A mRNA with two-step kinetics, while rd29B is induced only 3 h after the start of desiccation. The expression of both genes is stimulated about 3 h after application of ABA. It appears that rd29A has at least two cis-acting elements, one involved in the ABA-associated response to desiccation and the other induced by changes in osmotic potential. The -glucuronidase (GUS) reporter gene driven by the rd29A promoter was induced at significant levels by desiccation, cold, high-salt conditions and ABA in both transgenic Arabidopsis and tobacco. Histochemical analysis of GUS activity revealed that the rd29A promoter functions in almost all the organs and tissues of vegetative plants during water deficiency. 相似文献