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Query: EC:3.2.1.31 (
beta-glucuronidase
)
7,680
document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)
Genes encoding
phenylalanine ammonia-lyase
(
PAL
) form a small multigene family with at least three members in pea. Tissue-specific expression of the promoter of a member of
PAL
gene family (PSPAL1) was investigated in the transgenic tobacco transformants carrying the different modes of chimeric fusion between the PSPAL1 promoter and a bacterial
beta-glucuronidase
(GUS) gene. In stems, at least, strict correlation was found between steady-state levels of Gus-mRNA and enzyme activity. Significantly high level of GUS activity was observed in roots, particularly in meristematic tissues and the pigmented region of petals of transgenic tobacco carrying the translational fusion type B (-1,394 to +140 of PSPAL1 connected to Gus), followed by moderately high level of GUS activity carrying the translational fusion type A (-1,394 to +117). GUS expression in tissues of mature leaves, however, was very low in these constructs. Extremely low GUS activity was observed in the transformants of transcriptional fusion type (-1,394 to +5), whilst no activity was detected carrying non-transcription fusion type (-1,394 to -27). Furthermore, the pattern of the PSPAL1 expression was characterized in response to pathogen ingress and woundings in transgenic tobacco carrying the translational fusion type B. Woundings itself triggered marked expression of PSPAL1-driven GUS expression at the wounded sites. Inoculation of nonpathogens, Phytophthora capsici, P. boehmeriae and Erisiphe graminis f. sp. hordei, both caused rapid and very clear GUS expression zone along with the development of hypersensitive cell death area where callose was accumulated, however, the inoculation of a pathogen, P.nicotiana caused slow and hazy GUS expression zone along with the lesion development. These results suggest that the expression of pea PSPAL1 promoter is regulated in a similar fashion, at least in a part, in pea and transgenic tobacco, under the plant development and various environmental cues.
...
PMID:Temporal and spatial pattern of expression of the pea phenylalanine ammonia-lyase gene1 promoter in transgenic tobacco. 929 45
A study of the expression of a bean
phenylalanine ammonia-lyase
(
PAL
) promoter/
beta-glucuronidase
gene fusion in transgenic tobacco has shown that the PAL2 promoter has a modular organization. Expression of the PAL2 promoter in the vascular system involves positive and negative regulatory cis elements. Among these elements is an AC-rich motif implicated in xylem expression and a suppressing cis element for phloem expression. Using radiolabelled complementary oligonucleotides bearing the AC-rich motif, a cDNA clone encoding a DNA-binding protein has been isolated from a tobacco lambda gt11 expression library. This factor, named AC-rich binding factor (ACBF), showed binding specificity to the AC-rich region. The specificity of ACBF for the AC-rich region was also shown using a gel retardation assay with an ACBF recombinant protein extract. The deduced amino acid sequence from ACBF contains a long repeat of glutamine residues as found in well characterized transcription factors. Interestingly, ACBF shared sequence similarity to conserved amino acid motifs found in RNA-binding proteins. Genomic gel blot analysis indicated the presence of a small gene family of sequences related to ACBF within the tobacco nuclear genome. Analysis of tobacco mRNA using the ACBF cDNA as probe showed that while ACBF mRNA was present in all tissues examined, the highest transcript accumulation occurred in stem tissues. The functional characteristics of the AC-rich sequence were examined in transgenic tobacco. A heptamer of the AC-rich sequence, in front of a minimal 35S promoter from cauliflower mosaic virus (-46 to +4), conferred specific expression in xylem.
...
PMID:Characterization of a gene encoding a DNA-binding protein that interacts in vitro with vascular specific cis elements of the phenylalanine ammonia-lyase promoter. 934 52
To determine the regulatory mechanism of gene expression in the early stages of tracheary element (TE) differentiation, we isolated and characterized a genomic fragment of TED3 whose mRNA is expressed preferentially in differentiating TEs 12-24 h before morphological changes in the in vitro Zinnia system. Transgenic Arabidopsis plants with a chimeric gene of the 537 bp TED3 promoter and the
beta-glucuronidase
(GUS) reporter gene indicated the strong expression of the GUS gene by the TED3 promoter in TEs, in particular in immature TEs as well as stipules and trichomes. GUS expression driven by the promoter was also induced in callus, in which GUS activity was localized in immature TEs and slender cells around TEs that may be TE precursor cells. The TED3 promoter was not significantly activated by wounding. This pattern of expression differed clearly from that of other vascular tissue-related genes such as
PAL
, 4CL, and GRP1.8. The nature of TED3 promoter enables us to use it to monitor TE differentiation in tissue and to introduce foreign genes preferentially into immature TE.
...
PMID:Expression of the Zinnia TED3 promoter in developing tracheary elements of transgenic Arabidopsis. 952 Feb 82
The AC-rich motif, Pal-box, is an important cis-acting element for gene expression involved in phenylpropanoid biosynthesis. A cDNA clone (Ntlim1) encoding a Pal-box binding protein was isolated by Southwestern screening. The deduced amino acid sequence is highly similar to the members of the LIM protein family that contain a zinc finger motif. Moreover, Ntlim1 had a specific DNA binding ability and transiently activated the transcription of a
beta-glucuronidase
reporter gene driven by the Pal-box sequence in tobacco protoplasts. The transgenic tobacco plants with antisense Ntlim1 showed low levels of transcripts from some key phenylpropanoid pathway genes such as
phenylalanine ammonia-lyase
, hydroxycinnamate CoA ligase and cinnamyl alcohol dehydrogenase. Furthermore, a 27% reduction of lignin content was observed in the transgenic tobacco with antisense Ntlim1.
...
PMID:Functional analysis of tobacco LIM protein Ntlim1 involved in lignin biosynthesis. 1084 46
Lignin is a complex phenolic plant polymer that is essential for mechanical support, defense, and water transport in higher plants. The AC-rich motif, Pal-box is an important cis-acting element for gene expression in phenylpropanoid biosynthesis. We isolated a cDNA clone (Ntlim1) encoding a Pal-box binding protein by Southwestern screening. The deduced amino acid sequence of Ntlim1 is highly similar to members of the LIM protein family that contain a zinc finger motif. Moreover, Ntlim1 had a specific DNA-binding ability and transiently activated transcription of a
beta-glucuronidase
reporter gene driven by the Pal-box sequence. The results of transient expression assays with tobacco cultured cells showed that fusion proteins between GFP and Ntlim1 can enter nuclei. Transgenic tobacco plants with antisense Ntlim1 showed low levels of transcripts from some key phenylpropanoid pathway genes such as
phenylalanine ammonia-lyase
, hydroxycinnamate CoA ligase and cinnamyl alcohol dehydrogenase. Furthermore, a greater than 20% reduction in lignin content was observed in transgenic tobacco with antisense Ntlim1.
...
PMID:Transcriptional control of lignin biosynthesis by tobacco LIM protein. 1143 Sep 87
To elucidate heterologous promoter function in gymnosperms, we introduced the bean
phenylalanine ammonia-lyase
-
beta-glucuronidase
(PAL2-GUS) gene fusion into white pine (Pinus strobus L.). Over 15 lines were produced and integration of Agrobacterium T-DNA was confirmed by Southern analysis. Induction of the reporter gene was detected in all of the lines tested following UV illumination. In contrast, a weak but constant induction was seen in only a few lines following treatment with salicylic acid (SA) or jasmonic acid (JA). However, pretreatment of suspension cultures with SA or JA enhanced the induction of PAL2-GUS expression by UV irradiation. This specific enhancement or potentiation was reduced by 50% by treating the cells with indomethacin, an inhibitor of phospholipase activity, suggesting that the observed potentiation of UV induction involves the octadecanoid pathway. The UV induction was completely abolished by treating the cells with okadaic acid, an inhibitor of phosphatase activity. Thus, the induction of the heterologous PAL2 promoter from bean is consistent with the induction of
phenylalanine ammonia-lyase
(
PAL
) in angiosperms. Furthermore, our findings suggest that conifers, although phylogenetically distant to angiosperms, share some conserved promoter elements and some signal transduction mechanisms for UV-light perception.
...
PMID:Inducible expression of the heterologous PAL2 promoter from bean in white pine (Pinus strobus) transgenic cells. 1144 95
To understand molecular mechanisms underlying wound-induced expression of plant peroxidase genes, the promoter of a horseradish C2 peroxidase (prxC2) gene was analyzed. We had previously isolated a tobacco nuclear protein, Ntlim1, as a trans factor binding to a
PAL
-box motif of the prxC2 promoter; however, the function of the Ntlim1 trans factor and the
PAL
-box motif in wound-responsive expression of the prxC2 gene remains unclear. Here, we found that the prxC2 promoter without the intact
PAL
-box motif failed to direct a normal level of both the basal and the wound-induced expression of
beta-glucuronidase
(GUS) reporter gene in transgenic tobacco plants, indicating that the
PAL
-box motif functions as an essential cis element of the prxC2 promoter. We also found that antisense expression of Ntlim1 in transgenic plants carrying the prxC2 promoter::GUS chimeric construct decreased not only the level of the basal and the wound-induced expression of the GUS reporter gene but also the extent of wound inducibility of the prxC2 promoter itself. This result indicates that Ntlim1 is required for the basal level of prxC2 promoter activity as well as its up-regulation under wound stress. Moreover, consistent with the results obtained in planta, result from super-shift assay indicates that the Ntlim1 binds to the
PAL
-box motif independently of wound stress.
...
PMID:Ntlim1, a PAL-box binding factor, controls promoter activity of the horseradish wound-inducible peroxidase gene. 1208 67
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