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

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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)

A genomic clone of an acyl carrier protein gene (Bcg4-4) which is highly expressed in developing embryos of Brassica rapa was isolated and sequenced. The promoter and transcription terminator regions of Bcg4-4 were used to express a beta-glucuronidase reporter gene in transgenic rapeseed. Deletion of repeated domains in the promoter region did not lower beta-glucuronidase expression in seeds.
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PMID:Non-essential repeats in the promoter region of a Brassica rapa acyl carrier protein gene expressed in developing embryos. 153 31

A full-length cDNA clone encoding cytosolic glutamine synthetase (GS), expressed in roots and root nodules of soybean, was isolated by direct complementation of an Escherichia coli gln A- mutant. This sequence is induced in roots by the availability of ammonia. A 3.5-kilobase promoter fragment of a genomic clone (lambda GS15) corresponding to this cDNA was isolated and fused with a reporter [beta-glucuronidase (GUS)] gene. The GS-GUS fusion was introduced into a legume (Lotus corniculatus) and a nonlegume (tobacco) plant by way of Agrobacterium-mediated transformations. This chimeric gene was found to be expressed in a root-specific manner in both tobacco and L. corniculatus, the expression being restricted to the growing root apices and the vascular bundles of the mature root. Treatment with ammonia increased the expression of this chimeric gene in the legume background (i.e., L. corniculatus); however, no induction was observed in tobacco roots. Histochemical localization of GUS activity in ammonia-treated transgenic L. corniculatus roots showed a uniform distribution across all cell types. These data suggest that the tissue specificity of the soybean cytosolic GS gene is conserved in both tobacco and L. corniculatus; however, in the latter case, this gene is ammonia inducible. Furthermore, the ammonia-enhanced GS gene expression in L. corniculatus is due to an increase in transcription. That this gene is directly regulated by externally supplied or symbiotically fixed nitrogen is also evident from the expression of GS-GUS in the infection zone, including the uninfected cells, and the inner cortex of transgenic L. corniculatus nodules, where a flux of ammonia is encountered by this tissue. The lack of expression of GS-GUS in the outer cortex of the nodules suggests that ammonia may not be able to diffuse outside the endodermis.
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PMID:Ammonia-regulated expression of a soybean gene encoding cytosolic glutamine synthetase in transgenic Lotus corniculatus. 168 99

The requirements for homologous recombination between plasmid DNA molecules have been studied using the PEG (polyethylene glycol)-mediated transformation system of maize (Zea mays L.) protoplasts coupled with the transient expression assay for beta-glucuronidase (GUS). Two plasmids were introduced into maize protoplasts; one plasmid (pB x 26) contained a genomic clone of the Adh1 maize gene; the other plasmid (piGUS) was a promoterless construction containing part of intron A of the Adh1 gene fused to the gusA coding sequence. Thus, the two vectors shared an effective homologous region consisting of a 459 bp (HindIII-PvuII) fragment of the Adh1 intron A sequence. An active gusA fusion gene would result upon homologous recombination between the plasmids within the intron A sequence, and indeed GUS activity was observed in extracts following co-transformation of maize protoplasts with the two plasmids. The presence of recombinant DNA molecules in protoplast DNA isolated 1 day after co-transformation was verified using polymerase chain reactions (PCR) and Southern blots. For efficient homologous recombination, both plasmids had to be linearized. The recombination reaction was induced by restriction of the plasmid molecules either inside the effective homologous region or at the borders of the intron sequence. However, the presence of even small, terminal, nonhomologous sequences at the 3' end of the pB x 26 fragment inhibited the recombination reaction. Also, both ends of the linearized piGUS DNA molecules were involved in the recombination reaction. The results revealed some features of homologous recombination reactions occurring in plant cells which cannot be accommodated by mechanisms postulated for similar reactions in animal system and in lower eukaryotes.
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PMID:Homologous recombination between plasmid DNA molecules in maize protoplasts. 174 30

The developmental regulation of the translational elongation factor EF-1 alpha has been analyzed in tobacco. A gene fusion was constructed consisting of the 5' and 3' regions of the tomato genomic clone LeEF-A from the EF-1 alpha gene family and the beta-glucuronidase coding region. Analysis of the transgenic plants containing this chimeric gene demonstrated that the tomato LeEF-A flanking sequences were sufficient to confer expression patterns similar to those of the endogenous tobacco EF-1 alpha gene. The patterns of beta-glucuronidase activity in this system indicated that during plant growth and development EF-1 alpha is regulated with increased expression corresponding to regions of high protein synthesis, including meristems, rapidly growing tissues, and developing gametophytes. In addition, EF-1 alpha expression responds rapidly to changes in growth patterns induced by hormone treatment. Our results are in agreement with studies in animals indicating that EF-1 alpha expression may be rate limiting for protein synthesis and demonstrate that the analysis of EF-1 alpha is of value for studying interrelationships between protein synthesis and developmental control.
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PMID:Developmental analysis of elongation factor-1 alpha expression in transgenic tobacco. 184 19

We have isolated a putative transcription factor gene, PosF21, from Arabidopsis thaliana using an indirect cross-hybridization approach. cDNA clones were isolated which encode single repeating amino acids. Such sequences may function as activation domains in transcription factors and may be indicative for such proteins. The clone PosF21 encodes a region very rich in glutamines. Besides this putative activation domain it encodes a protein sequence which shows all the characteristics of a basic-domain/leucine zipper type of DNA-binding domain. PosF21 is expressed constitutively at a low level in young seedlings and in roots, stems and leaves of mature Arabidopsis plants. A genomic clone of PosF21 was isolated and the gene structure was analyzed. Related sequences in Arabidopsis and a wide range of other plants were detected using the putative DNA-binding domain as a probe in cross-hybridization experiments. Transient transformations in tobacco protoplasts were performed using the beta-glucuronidase (GUS) gene as reporter gene. Approximately 400 bp of the 5' genomic region of PosF21 promote expression of the GUS gene in tobacco protoplasts. Evidence for a regulatory function of PosF21 was obtained since co-expression of the full PosF21 protein or its DNA-binding region alone specifically stimulated GUS gene expression directed from the PosF21 promoter by 6-8-fold.
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PMID:Isolation and molecular characterization of PosF21, an Arabidopsis thaliana gene which shows characteristics of a b-Zip class transcription factor. 184 85

We have isolated and characterized the genomic clone lambda CHN50 corresponding to tobacco basic endochitinase (E.C.3.2.1.14). DNA sequence and blotting analysis reveal that the coding sequence of the gene present on lambda CHN50 is identical to that of the cDNA clone pCHN50 and, moreover, the CHN50 gene has its origin in the progenitor of tobacco, Nicotiana sylvestris. Tobacco basic chitinases are encoded by a small gene family that consists of at least two members, the CHN50 gene and a closely related CHN17 gene which was characterized previously. By northern blot analysis, it is shown that the CHN50 gene is highly expressed in suspension-cultured tobacco cells and the mRNA accumulates at late logarithmic growth phase. To identify cis-DNA elements involved in the expression of the CHN50 gene in suspension-cultured cells, the chimeric gene consisting of 1.1 kb CHN50 5' upstream region fused to the coding sequence of beta-glucuronidase (GUS) was introduced by electroporation into protoplasts isolated from suspension-cultured tobacco cells. Transient GUS activity was found to be dependent on the growth phase of the cultured cells, from which protoplasts had been prepared. Functional analysis of 5' deletions suggests that the distal region between -788 and -345 contains sequences that potentiate the high-level expression in tobacco protoplasts and the region (-68 to -47) proximal to the TATA box functions as a putative silencer.
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PMID:Gene structure and expression of a tobacco endochitinase gene in suspension-cultured tobacco cells. 188 89

We investigated the 5'-upstream region of the gene encoding the catalytic subunit of the V-type H(+)-ATPase in Daucus carota. A genomic sublibrary was screened with a cDNA probe, and a 4-kilobase genomic clone was obtained covering the first two exons and about 3 kilobases of the 5'-upstream sequence. The intron/exon boundaries matched established consensus sequences. Within 240 base pairs (bp) upstream of the initiation codon three putative TATA boxes were found. Ribonuclease protection and primer extension analysis indicated that the three TATA boxes corresponded to two major and one minor transcription start sites. The flanking sequences of the two more proximal TATA boxes were nearly identical. Additional sequence motifs with putative regulatory function are two CCAAT boxes, an Sp1-binding consensus sequence, and long (TATA)n stretches within 800 bp of the 5'-upstream sequence. Transcriptional fusions to the beta-glucuronidase reporter gene were made for two different promoter constructs, and the resulting plasmids were mobilized into Agrobacterium tumefaciens. The analysis of beta-glucuronidase activities in the transformed carrot calli showed that 240 bp of the upstream sequence, including all three TATA boxes, led to low but detectable beta-glucuronidase expression; however, the larger construct, which included the putative Sp1-binding sequence and the (TATA)n stretches, led to an approximately 6-fold higher beta-glucuronidase expression. Histochemical analysis of beta-glucuronidase activity in the transformed calli showed no preferential expression in any specific cell type, in keeping with the presumed "housekeeping" character of the V-type H(+)-ATPase catalytic subunit gene.
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PMID:Structure and function of the promoter of the carrot V-type H(+)-ATPase catalytic subunit gene. 213 75

We have isolated a genomic clone of an auxin-regulated par gene, which is expressed during the transition from G0 phase to S phase in the early stage of tobacco mesophyll protoplasts cultured in vitro, from a tobacco genomic library using the par cDNA as a probe. When a chimeric gene, in which a reporter gene for bacterial beta-glucuronidase (GUS) was placed downstream of the 5' flanking sequences of the par gene, was introduced into tobacco mesophyll protoplasts by electroporation, the chimeric gene elicited auxin-regulated expression of GUS activity. Because deletion of a 111-base-pair (bp) direct repeat in the 5' flanking sequences of the par gene abolished the auxin-induced GUS activity, it is deduced that in the 111-bp direct repeat of the par gene promoter is localized an auxin-responsive region, which regulates auxin-mediated activation of transcription.
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PMID:Location of the cis-acting auxin-responsive region in the promoter of the par gene from tobacco mesophyll protoplasts. 223 15

A set of 5' promoter deletions from Zmg13, a genomic clone of a pollen-specific gene of maize, has been transcriptionally fused to a beta-glucuronidase (GUS) reporter gene in the binary vector pBI101. Tobacco leaf disks were transformed and mature plants analyzed for GUS activity directed by the Zmg13 promoter constructs. Transgenic plants containing the 375 bp Zmg13 sequence from -314 to +61 relative to the transcription start site transcribed GUS RNA and expressed active GUS enzyme in mature pollen but not in leaves. Plants transformed with a 35S CaMV promoter-GUS transcriptional fusion expressed GUS RNA in leaves but not in pollen. Neither GUS RNA or active enzyme could be detected in pollen or leaves from plants containing a 124 bp Zmg13-GUS transcriptional fusion missing the putative Zmg13 TATA box. No GUS RNA or enzyme expression was not detected in non-transformed tobacco. RNA and GUS histochemical analysis of the T1 generation confirmed that the temporal expression pattern of Zmg13-GUS transcription in tobacco followed that of the native gene in maize and that the Zmg13 promoter sequences from the maize gene are able correctly to direct genetically stable, tissue-specific gene expression in transgenic tobacco plants.
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PMID:Promoter sequences from a maize pollen-specific gene direct tissue-specific transcription in tobacco. 227 35

The complete nucleotide sequence of murine beta-glucuronidase (GUS) mRNA has been compiled from three overlapping cloned cDNAs and a single GUS-specific genomic clone. The sequence is composed of 2455 nucleotides, exclusive of the poly(A) tail. The 5' and 3' untranslated regions contain 12 and 499 bases, respectively, with the open reading frame encoding a polypeptide of 648 amino acids (74.2 kDa), including a 22 amino acid signal sequence. The nucleotide and deduced amino acid sequences of murine GUS are compared to those published for rat and human GUS and the results are presented. Murine GUS also shares amino acid sequence identity with Escherichia coli GUS and beta-galactosidase. The complete sequences of murine GUS mRNA and its deduced polypeptide provide a basis from which to study the mechanisms responsible for the well-characterized variation in GUS expression among inbred mouse strains.
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PMID:The complete nucleotide sequence of murine beta-glucuronidase mRNA and its deduced polypeptide. 339 60


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