Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound

---

Query: EC:3.2.1.31 (beta-glucuronidase)
7,680 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

We have successfully transferred and expressed a reporter gene driven by an alpha-amylase promoter in a japonica type of rice (Oryza sativa L. cv. Tainung 62) using the Agrobacterium-mediated gene transfer system. Immature rice embryos (10-12 days after anthesis) were infected with an Agrobacterium strain carrying a plasmid containing chimeric genes of beta-glucuronidase (uidA) and neomycin phosphotransferase (nptII). Co-incubation of potato suspension culture (PSC) with the Agrobacterium inoculum significantly improved the transformation efficiency of rice. The uidA and nptII genes, which are under the control of promoters of a rice alpha-amylase gene (alpha Amy8) and Agrobacterium nopaline synthase gene (nos), respectively, were both expressed in G418-resistant calli and transgenic plants. Integration of foreign genes into the genomes of transgenic plants was confirmed by Southern blot analysis. Histochemical localization of GUS activity in one transgenic plant (R0) revealed that the rice alpha-amylase promoter functions in all cell types of the mature leaves, stems, sheaths and roots, but not in the very young leaves. This transgenic plant grew more slowly and produced less seeds than the wild-type plant, but its R1 and R2 progenies grew normally and produced as much seeds as the wild-type plant. Inheritance of foreign genes to the progenies was also confirmed by Southern blot analysis. These data demonstrate successful gene transfer and sexual inheritance of the chimeric genes.
...
PMID:Agrobacterium-mediated production of transgenic rice plants expressing a chimeric alpha-amylase promoter/beta-glucuronidase gene. 839 95

DNA fragments from the bidirectional promoter region of the geminivirus chloris striate mosaic virus (CSMV) were cloned into the pUC18-based vector, pG1 producing transcriptional fusions with the beta-glucuronidase (GUS) gene and nopaline synthase terminator sequence. The relative activity of each promoter construct was analyzed by a GUS expression assay of extracts from Zea mays (maize) Black Mexican Sweet protoplasts coelectroporated with the GUS reporter constructs and constructs in which individual CSMV open reading frames (ORFs) were placed under control of a cauliflower mosaic virus 35 S promoter. Weak promoter activity was observed for the promoter of the C1 and C2 ORFs (C1-C2 gene) and for the promoter of the V1 ORF. The activity of these promoters was unaffected by coelectroporation with the CSMV ORF constructs. Moderate activity was observed for the promoter of the V2 ORF (coat protein gene) which was enhanced by coelectroporation of the C1-C2 ORF construct. Sequences within the C1-C2 gene responsible for transactivation of the V2 ORF promoter were mapped close to the A site of a conserved NTP-binding sequence pattern within the C2 ORF. To a lesser extent activity for the promoter of the V2 ORF was enhanced by the V2 ORF construct providing evidence for positive autoregulation of the CSMV coat protein gene.
...
PMID:The activity of the coat protein promoter of chloris striate mosaic virus is enhanced by its own and C1-C2 gene products. 843 84

A series of chimeric promoters for higher-level expression of foreign genes in plants was constructed as fusions of a gene for beta-glucuronidase (GUS) with the terminator of a gene for nopaline synthase (nos) or of the cauliflower mosaic virus (CaMV) 35S transcript, and the strength of these promoters was assayed in transient and stable expression systems in tobacco and rice. As parts of these promoters, the CaMV 35S core promoter, three different 5'-upstream sequences of the 35S promoter, the first intron of a gene for phaseolin, and a 5'-untranslated sequence (omega sequence) of tobacco mosaic virus were used in various combinations. In tobacco and rice protoplasts, all three fragments of the 35S promoter (-419 to -90, -390 to -90 and -290 to -90, relative to the site of initiation of transcription), the intron, and the omega sequence effectively enhanced GUS activity. Some chimeric promoters allowed levels of GUS activity that were 20- to 70-fold higher than those obtained with the 35S promoter in pBI221. In tobacco protoplasts, the two longer fragments of the 35S promoter were more effective than the shortest fragment. In rice cells, by contrast, the shortest fragment was as effective as the two longer ones. The terminator of the 35S transcript was more effective than that of the nos gene for gene expression. In transgenic tobacco plants, a representative powerful promoter, as compared to the 35S promoter, allowed 10- and 50-fold higher levels of expression on average and at most, respectively, with no clear qualitative differences in tissue- and organ-specific patterns of expression. When the representative promoter was introduced into tobacco with a gene for luciferase, the autofluorescence of detached leaves after a supply of luciferin to petioles was great and was easily detectable by the naked eye in a dark room.
...
PMID:Efficient promoter cassettes for enhanced expression of foreign genes in dicotyledonous and monocotyledonous plants. 872 Sep 24

Transcriptionally regulated expression of tobacco anionic peroxidase was investigated with regard to tissue specificity and developmental regulation. Two tobacco species, Nicotiana sylvestris and Nicotiana tabacum cv. Xanthi, were stably transformed with a gene chimera composed of 3 kb of the tobacco anionic peroxidase promoter, the Escherichia coli beta-glucuronidase (GUS) coding region and the nopaline synthase terminator. Gene expression was regulated spatially and developmentally in all organs, and generally increased with age and maturity of the plant, tissue or organ. In the aerial portions of the plant, GUS activity was strongly expressed in trichomes and epidermis at nearly all developmental stages. In later stages of development, activity was also detected in ground tissue and parenchyma cells associated with vascular tissues. Activity in roots was limited to cortical cells and vascular-associated parenchyma cells. In reproductive tissue, expression was observed in sepals and petals before anthesis, and in all floral organs after anthesis. Expression was never detected in vascular tissue and was poorly correlated with lignification except in the cells surrounding primary xylem and pericyclic fibers in N. sylvestris. These studies suggest that this peroxidase isoenzyme is only limitedly involved in lignification but may be important in plant defense, growth and development.
...
PMID:Expression of the tobacco anionic peroxidase gene is tissue-specific and developmentally regulated. 948 46

The identification of regulatory elements conferring high levels of expression in differentiating pine xylem will be valuable for genetic engineering of wood properties and will contribute to our understanding of gene regulation in this important group of forest trees. We examined the roles of both upstream and downstream elements in regulating the expression of two genes with preferential expression in developing xylem of loblolly pine. Gene constructs containing a PtX3H6, PtX14A9, or CaMV 35S promoter, the uidA gene encoding beta-glucuronidase, and a PtX3H6, PtX14A9, or NOS terminator were used to transform tobacco and hybrid poplar. When combined with the NOS terminator, neither pine promoter conferred xylem-specific expression in tobacco. When combined with the PtX3H6 promoter, an element at the 3' end of PtX3H6 reduced GUS expression resulting in preferential expression in vascular tissues. This silencing effect was not observed when the pine terminator was tested in conjunction with the CaMV 35S promoter. The PtX14A9 terminator did not increase tissue specificity. In leaves of transgenic poplar, both pine promoters conferred preferential GUS expression in veins when combined with the NOS terminator. The PtX3H6 terminator greatly decreased expression in leaves and stems when combined with the PtX3H6 promoter but only slightly altered expression when combined with the CaMV 35S promoter. An element at the 3' end of PtX14A9 increased GUS expression in veins when used in conjunction with either the PtX14A9 or CaMV35S promoter.
...
PMID:Sequences upstream and downstream of two xylem-specific pine genes influence their expression. 1116 79

The soybean tubB1 gene is expressed primarily in the germinating seedling and is strongly down regulated in response to light in the upper hypocotyl. Previous studies demonstrate that the 1 kb 5'-flanking region of this gene is sufficient for its appropriate expression in etiolated seedlings. Transient expression studies demonstrated that the presence of the tubB1 3'-flanking sequence element decreased reporter gene expression as compared to the nopaline synthase (NOS) 3'-flanking sequence element. In this study we investigated the ability of the 3' flanking region to influence the expression of a beta-glucuronidase (GUS) reporter gene in transgenic tobacco and Arabidopsis. The presence of the tubB1 3'-flanking sequence element in chimera constructs reduced reporter gene expression specifically in the hypocotyl and petioles of light-grown, transgenic seedlings. Additionally, site-directed mutagenesis of the two TATA sequences in the 1 kb tubB1 5'-flanking sequence element (TATA box A in the -122 to -117 bp region and TATA box B in the -35 to -30 region) showed that both elements are functional and additive in controlling tubB1 gene expression in seedling tissues. While transcription from TATA box A was predominant regardless of lighting conditions, the relative usage of TATA box B increased in the dark. We conclude that both TATA box sequences are utilized to direct expression of the tubB1 gene to the cotyledons, hypocotyl and root tip of germinating seedlings that are regions of cell expansion and that the 3'-flanking sequence element down-regulates expression in the hypocotyl in response to light. Thus, it is plausible that the tubB1 protein may play an important role in cell expansion in seedling development requiring its regulated expression by light.
...
PMID:The roles of two TATA boxes and 3'-flanking region of soybean beta-tubulin gene (tubB1) in light-sensitive expression. 1171 May 21

Mature zygotic embryos of loblolly pine (Pinus taeda L.) were transformed by Agrobacterium tumefaciens strain LBA 4404 harbouring the plasmid pBI121 which carried the selectable marker gene, neomycin phosphotransferase II (npt II) controlled by the promoter of the nopaline synthase gene, and the uidA reporter gene, encoding beta-glucuronidase (GUS) driven by the cauliflower mosaic virus 35S promoter. Organogenic transgenic calli and transgenic regenerated plantlets were produced on selection medium containing 15 mg/L kanamycin, and confirmed by GUS histochemical staining, polymerase chain reaction (PCR), and southern blot analysis. Influences of phytohormone (BA/IBA) and antibiotics on growth and differentiation of organogenic transgenic calli were investigated. Of the phytohormone (BA/IBA) and antibiotics administered, 500 mg/L carbenicillin combined with 2 mg/L BA and 0.5 mg/L IBA (BA/IBA = 4) resulted in a 54.2% higher increase in the growth of transgenic calli as well as in the differentiation of transgenic calli, which was 45.7% more than that of control on the 6th week of culture. Claforan at 500 mg/L combined with 2 mg/L BA and 0.5 mg/L IBA resulted in a 40.8% increase in the growth of transgenic calli, and 38.7% increase in the frequency of transgenic calli forming adventitious shoots compared with the control. The growth and differentiation of transgenic calli of loblolly pine was reduced preferentially by higher BA/IBA (BA/IBA = 8), as well as high concentration of antibiotics (carbenicillin and claforan, 550 mg/L each). But it was observed that 450 mg/L and 500 mg/L carbenicillin and claforan caused an increase in growth and differentiation of transgenic calli. These results suggested that the establishment of an efficient Agrobacterium tumefaciens-mediated transformation protocol for stable integration of foreign genes into loblolly pine was also dependent on the regulation of phytohormone and antibiotic on growth and differentiation of transgenic calli. This work could be useful for the future studies of genetic transformation of conifers.
...
PMID:Growth and differentiation of transgenic callus regulated by phytohormones and antibiotics in transformation of loblolly pine. 1190 1

One of the strategies to make crops resistant to the beet cyst nematode Heterodera schachtii is the destruction of the feeding site or syncytium. This can be achieved by local expression of the cytotoxic barnase gene under control of a nematode-inducible plant promoter that is active in the syncytium. Expression of barnase outside the feeding site has to be neutralized by its inhibitor barstar driven from a constitutive promoter that is downregulated in the syncytium. Several promoters that are upregulated in feeding structures were identified using the promoter tagging strategy in Arabidopsis thaliana (Barthels et al., 1997) or by differential cDNA screening in tomato (Van der Eycken et al., 1996). Nematode downregulated promoters in Arabidopsis were described by Goddijn et al. (1993). Five nematode-induced promoters (ARM1, 1164, 728, 25 and Lemmi9) and four downregulated promoters (CaMV35S, the nopaline synthase promoter (nos) and the rooting loci promoters RolC and RolD) fused to the beta-glucuronidase (gus) reporter gene were introduced into sugar beet hairy roots by transformation with Agrobacterium rhizogenes to evaluate their expression pattern. All upregulated promoters were found to be active at the base of lateral roots. The 728 and 25 promoter were as well active in root tips. In the 25-gus lines GUS could also be detected in the vascular tissue, while the ARM1 promoter was also active in sugar beet callus. The Lemmi9 promoter and the 4 constitutive promoters were active in the entire root. The transgenic hairy roots were inoculated with Heterodera schachtii and at different time-points (4, 8, 15, 22 days after inoculation; dpi) GUS analysis was performed on the infected roots. For the ARM1, 1164 and 728 promoter the highest gus expression level in syncytia was observed at 8 dpi. In 4 days old syncytia of the 25-gus lines the intensity of the GUS signal was of the same extent as the non-specific vascular signal. In later stages it even disappeared from the feeding sites. The gus expression level in syncytia of Lemmi9-gus hairy roots was equal to that in control roots. The RolC and 35S promoter were found to be downregulated at 8 dpi, the RolD promoter at 15 dpi and the nos promoter already at 4 dpi.
...
PMID:Analysis of nematode-responsive promoters in sugar beet hairy roots. 1242 82

Fruit ripening is associated with a number of physiological and biochemical changes. They include degradation of chlorophyll, synthesis of flavor compounds, carotenoid biosynthesis, conversion of starch to sugars, cell wall solublisation and fruit softening. These changes are brought about by the expression of specific genes. People are interested in the molecular mechanism involved in the regulation of gene transcription during fruit ripening. Many fruit-specific promoters such as PG, E4, E8, and 2A11 have been characterized and shown to direct ripening-specific expression of reporter genes. AGPase plays the key role in catalyzing the biosynthesis of starch in plants. It is a heterotetrameric enzyme with two small subunits and two large subunits, which are encoded by different genes. In higher plants, small subunits are highly conserved among plant species and expressed in all tissues. And the large subunits are present at multiple isoforms and expressed in a tissue-specific pattern. In fruits, the expression pattern of the large subunits varies with plant species. That made it important to study the transcriptional regulation of the large subunits of AGPase in different plant species. Northern-blot analysis indicates in watermelon, an isoform of the large subunits Wml1 expressed specifically in fruits, not in leaves. The 5' flanking region of Wml1, which covers 1573bp, has been isolated through the method of uneven PCR. And transient expression assay has shown that the 1573bp (named WSP) can direct fruit-specific expression of GUS gene. Our goal in this study was to scan the promoter region for main regulatory regions involved in fruit-specific expression. A chimaeric gene was constructed containing the WSP promoter, the beta-glucuronidase (GUS) structural sequence as a reporter gene and the nopaline synthase polyadenylation site (NOS-ter). The plasmid pSPA was digested with Hind III + Hinc II and promoter fragment of 1573bp (from 180bp to 1752bp) was cut out and cloned into Sma I sites of pBluescript SK(-), to produce pBSPA-16. The same insert was then cut out with Hind III + BamH I, and ligated with transient expression vector pBI426 digested by HindIII + Bgl II to produce pISPA-16. Three 5'-end deletions of the promoter were obtained and fused to GUS gene in plant transient expression vector pBI426: the 1201bp fragment (from 551bp to 1752bp) was generated by digestion of pBSPA-16 with BamH I + SnaB I, the 898bp fragment (from 854bp to 1752bp) by BamH I + EcoRV. Both fragments were ligated with pBluescript SK(-) digested by BamH I + Sma I, to produce pBSPA-12 and pBS-PA-9. The inserts were cut out with HindmIII + BamH I and ligated with pBI426 digested by Hind III + Bgl II, to produce pISPA-12 and pISPA-9. The 795bp fragment (from 957bp to 1752bp) was generated by digestion of pSPA with Hinc II + EcoR I, promoter fragment was cut out and cloned into Sma I sites of pBluescript SK(-), to produce pBSPA-8. The same insert were cut out with Hind III + BamH I, and ligated with transient expression vector pBI426 digested by Hind III + Bgl II. The 1573bp fragment and three 5'-end deletions were delivered into watermelon leaf, stem, flower and fruit of different development stages (5, 10, 20 days after pollination) via particle bombardment using a biolistic PDS-1000/He particle gun. Bombardment parameters were as follows: a helium pressure of 1200 psi, vacuum of 91432.23Pa, 7 cm between the stopping screen and the plate. Histochemical assay were done on all the tissues bombarded after incubation for 2 days. The 1573bp fragment had the strongest promoter activity, and can induce GUS expression in fruits of 5 and 20 days after anthesis and flowers, but not in fruits of 10 days after anthesis, leaves and stems. Fragments of 1201bp and 898bp can induce GUS expression only in fruits of 20 days after anthesis, and with lower expression levels than 1573bp. Fragment of 795bp was not able to direct GUS expression in any of the tissues bombarded (data not shown). It can be concluded that of the 1573bp, 1201 bp, 898bp Wml1 5'flanking regions include the necessary information directing fruit-specific expression. Deletion from 180bp to 551bp doesn't affect the fruit-specificity of the promoter, but lowered the expression level. There may be some cis-acting elements located in this region, which can enhance external gene expression in later stages of fruit development. Deletion from 854bp and 958bp led to loss of GUS expression. This region includes the necessary information needed for gene expression as well as the regulatory elements for fruit-specific transcription.
...
PMID:[Primary targeting of functional regions involved in transcriptional regulation on watermelon fruit-specific promoter WSP]. 1596 27

Agrobacterium tumefaciens is established as a vector for gene transfer in many dicotyledonous plants but is not accepted as a vector in monocotyledonous plants, especially in the important Gramineae. The use of Agrobacterium to transfer genes into monocot species could simplify the transformation and improvement of important crop plants. In this report we describe the use of Agrobacterium to transfer a gene into corn, the regeneration of plants, and detection of the transferred genes in the F(1) progeny. Shoot apices of Zea mays L. variety Funk's G90 were cocultivated with A. tumefaciens EHA 1, which harbored the plasmid pGUS3 containing genes for kanamycin resistance (NPT II) and beta-glucuronidase (GUS). Plants developed from these explants within 4 to 6 weeks. Fluorometric GUS assays of leaves and immature seeds from the plants exhibited low GUS activity. Both NOS and GUS gene fragments were amplified by polymerase chain reaction in the DNA isolated from the F(1) generations of one of the original transformed plants. Southern analysis showed both GUS and NPT probes hybridized to DNA in several of the F(1) progeny, demonstrating the incorporation of GUS and NPT II genes into high molecular weight DNA. These data establish successful gene transfer and sexual inheritance of the genes.
...
PMID:Transformation of Zea mays L. Using Agrobacterium tumefaciens and the Shoot Apex. 1666 1


<< Previous 1 2 3 Next >>

---