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1.
A 577-bp promoter segment of Agrobacterium rhizogenes rolC, previously known as the phloem-specific gene expression promoter, was fused to the 5′ end of a reporter gene, β-glucuronidase
(GUS), uidA. This rolC-promoter-driven expression of the GUS gene was found to be significantly strong in glandular cells in transgenic tobacco plants. Analysis of this segment of the
promoter sequence revealed a myb response element. 相似文献
2.
The Perilla (Perilla frutescens L. cv. Okdong) oleosin gene, PfOle19, produces a 19-kDa protein that is highly expressed only in seeds. The activity of the −2,015 bp 5′-upstream promoter region
of this gene was investigated in transgenic Arabidopsis plants using the fusion reporter constructs of enhanced green fluorescent protein (EGFP) and β-glucuronidase (GUS). The PfOle19 promoter directs Egfp expression in developing siliques, but not in leaves, stems or roots. In the transgenic Arabidopsis, EGFP fluorescence and histochemical GUS staining were restricted to early seedlings, indehiscent siliques and mature seeds.
Progressive 5′-deletions up to the −963 bp position of the PfOle19 promoter increases the spatial control of the gene expression in seeds, but reduces its quantitative levels of expression.
Moreover, the activity of the PfOle19 promoter in mature seeds is 4- and 5-fold greater than that of the cauliflower mosaic virus 35S promoter in terms of both
EGFP intensity and fluorometric GUS activity, respectively. 相似文献
3.
Xiaobo Qin Xiaojiang Zheng Caixia Shao Jihai Gao Luding Jiang Xunlu Zhu Fang Yan Lin Tang Ying Xu Fang Chen 《Planta》2009,230(2):387-395
Ribosome-inactivating proteins (RIPs) represent a type of protein that universally inactivates the ribosome thus inhibiting
protein biosynthesis. Curcin-L was a type I RIP found in Jatropha curcas L.. Its expression could be activated in leaves by treatments with abscisic acid, salicylic acid, polyethylene glycol, temperature
4, 45°C and ultraviolet light. A 654 bp fragment of a 5′ flanking region preceding the curcin-L gene, designated CP2, was
cloned from the J. curcas genome and its expression pattern was studied via the expression of the β-glucuronidase (GUS) gene in transgenic tobacco.
Analysis of GUS activities showed that the CP2 was leaf specific, and was able to drive the expression of the reporter gene
under stress-induction conditions. Analysis of a series of 5′-deletions of the CP2 suggested that several promoter motifs
were necessary to respond to environmental stresses. 相似文献
4.
The promoter of the pepper pathogen-induced membrane protein gene CaPIMP1 was analyzed by an Agrobacterium-mediated transient expression assay in tobacco leaves. Several stress-related cis-acting elements (GT-1, W-box and ABRE) are located within the CaPIMP1 promoter. In tobacco leaf tissues transiently transformed with a CaPIMP1 promoter-β-glucuronidase (GUS) gene fusion, serially 5′-deleted CaPIMP1 promoters were differentially activated by Pseudomonas syringae pv. tabaci, ethylene, methyl jasmonate, abscisic acid, and nitric oxide. The −1,193 bp region of the CaPIMP1 gene promoter sequence exhibited full promoter activity. The −417- and −593 bp promoter regions were sufficient for GUS gene activation by ethylene and methyl jasmonate treatments, respectively. However, CaPIMP1 promoter sequences longer than −793 bp were required for promoter activation by abscisic acid and sodium nitroprusside treatments.
CaPIMP1 expression was activated in pepper leaves by treatment with ethylene, methyl jasmonate, abscisic acid, β-amino-n-butyric acid, NaCl, mechanical wounding, and low temperature, but not with salicylic acid. Overexpression of CaPIMP1 in Arabidopsis conferred hypersensitivity to mannitol, NaCl, and ABA during seed germination but not during seedling development. In contrast,
transgenic plants overexpressing CaPIMP1 exhibited enhanced tolerance to oxidative stress induced by methyl viologen during germination and early seedling stages.
These results suggest that CaPIMP1 expression may alter responsiveness to environmental stress, as well as to pathogen infection.
The nucleotide sequence data reported here has been deposited in the GenBank database under the accession number DQ356279. 相似文献
5.
A 1535 bp promoter of the nitrate reductase gene (nia) from birch (Betula pendula) and a series of 5′ deletions were fused to the β-glucuronidase (GUS) gene and introduced into Nicotiana plumbaginifolia. In transgenic plants the NR promoter sequences directed strong GUS expression in the root epidermal hair cells, and in phloem
cells of leaf and stem vascular tissue. The NR promoter confers also a significant stimulation of the GUS gene expression
by nitrate. These findings might indicate that nitrate flow is one of the signals involved into tissue and cell specific expression
of the NR promoter GUS fusions.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
6.
The promoter of Brassica campestris Male Fertile 5 (BcMF5), a pollen coat protein member, class A (PCP-A) gene family, was isolated from Brassica rapa L. ssp. chinensis Makino (Chinese cabbage-pak-choi) by Thermal Asymmetric Interlaced Polymerase Chain Reaction (TAIL-PCR). Sequence analysis
suggested that the 605-bp promoter of BcMF5 appears to be a pollen promoter. In an attempt to confirm the promoter activity of BcMF5 promoter, −609 to +3 bp and −377 to +3 bp fragments of the upstream sequence of BcMF5 were inserted at the site upstream of the coding region of the uidA gene in the sense orientation to construct two deletion expression vectors. Transient expression analysis in onion epidermal
cells by particle bombardment showed that both −609 to +3 bp and −377 to +3 bp fragments of BcMF5 promoter were capable of driving β-glucuronidase gene expression. Furthermore, by Agrobacterium-mediated genetic transformation
method, Arabidopsis transgenic KanR plants were obtained. GUS assay analysis revealed that the promoter of BcMF5 induced gene expression at the early stage of anther development and drove high levels of GUS expression in anther walls,
upper regions of petals, pollen, and pollen tubes in the middle and late stage of anther development, but did not drive any
expression in sepals and pistils. 相似文献
7.
Ozone- and ethylene-induced regulation of a grapevine resveratrol synthase promoter in transgenic tobacco 总被引:1,自引:0,他引:1
Bernhard Grimmig Roland Schubert Regina Fischer Rüdiger Hain Peter H. Schreier Christian Betz Christian Langebartels Dieter Ernst Heinrich Sandermann 《Acta Physiologiae Plantarum》1997,19(4):467-474
Stilbene synthases (STSs) are enzymes that play a critical role in the biosynthesis of stilbene, phytoalexins in a small number
of unrelated plant species, and are induced by various biotic and abiotic stressors like pathogen attack, UV-irradiation or
ozone exposure. To investigate the molecular basis for ozone-induced plant stress responses, we have examined the promoter
of the grapevine resveratrol synthase (Vst1). In this report we summarize the influence of ozone on gene regulation. In transgenic tobacco a chimeric gene construct,
containing the Vst1 promoter combined with the β-glucuronidase (GUS) reporter gene, is rapidly induced by ozone (0.1 μl·l−1, 12 h). The same construct is also strongly induced by ethylene (20 μl·l−1, 12 h). Promoter deletion analysis of the 5′ flanking sequence identified a positive regulatory element between −430 bp and
−280 bp. This region contains ethylene-responsive enhancer elements, as well as an elicitor-responsive sequence in inverse
orientation. 相似文献
8.
Sergei F. Krasnyanski Jagdeep Sandhu Leslie L. Domier Dennis E. Buetow Schuyler S. Korban 《In vitro cellular & developmental biology. Plant》2001,37(4):427-433
Summary Two different promoters, a cauliflower mosaic virus (CaMV) 35S promoter with a 5′-untranslated leader sequence from alfalfa
mosaic virus RNA4 (designated as CaMV 35S/AMV) and an E-8 fruit-ripening-specific promoter, were compared to evaluate their
effects on expression of the uidA reporter gene in transgenic tomato plants. In order to generate sufficient numbers of transgenic tomato plants, both a reliable
regeneration system and an efficient Agrobacterium transformation protocol were developed using 8-d-old cotyledons of tomato (Lycopersicon ecsulentum Mill. cv. Swifty Belle). Two sets of constructs, both derivatives of the binary vector pBI121, were used in transformation
of tomato whereby the uidA gene was driven either by the CaMV 35S/AMV or the E-8 fruit-ripening-specific promoter. Southern blot hybridization confirmed
the stable integration of the chimeric uidA gene into the tomato genome. Fruit and leaf tissues were collected from T0 and T1 plants, and assayed for β-glucuronidase (GUS) enzyme activity. As expected, both vegetative and fruit tissues of transgenic
plants carrying the uidA gene under the control of CaMV 35S/AMV showed varying levels of GUS activity, while no expression was observed in vegetative
tissues of transgenic plants carrying the uidA gene driven by the E-8 promoter. All fruits from transgenic plants produced with both sets of constructs displayed expression
of the uidA gene. However, when this reporter gene was driven by the CaMV 35S/AMV, GUS activity levels were significantly higher than
when it was driven by the E-8 fruit-specific promoter. The presence/absence of the uidA gene in T1 plants segregated in a 3∶1 Mendelian ratio. 相似文献
9.
10.
Nancy L. Paiva Abraham Oommen Maria J. Harrison Richard A. Dixon 《Plant Cell, Tissue and Organ Culture》1994,38(2-3):213-220
Isoflavonoids are believed to play important roles in plant-microbe interactions. During infection of alfalfa (Medicago sativa) leaves with the fungal pathogen Phoma medicaginis, rapid increases in mRNA levels and enzyme activities of isoflavone reductase, phenylalanine ammonia-lyase, chalcone synthase and other defense genes are observed within 1 to 2 hours. The phytoalexin medicarpin and its antifungal metabolite sativan increase beginning at 4 and 8 hours, respectively, along with other isoflavonoids. In contrast, during colonization of alfalfa roots by the symbiotic mycorrhizal fungus Glomus versiforme, expression of the general phenylpropanoid and flavonoid genes phenylalanine ammonia-lyase and chalcone synthase increases while mRNA levels for the phytoalexin-specific isoflavone reductase decrease. The total isoflavonoid content of colonized roots increases with time and is higher than that of uninoculated roots, but the accumulation of the antifungal medicarpin is somehow suppressed.An isoflavone reductase genomic clone has been isolated, promoter regions have been fused to the reporter gene -glucuronidase, and the promoter-reporter fusions have been transformed into tobacco and alfalfa. Using histological staining, we have studied the developmental and stress-induced expression of this phytoalexin-specific gene in whole plants at a more detailed level than other methods allow. The isoflavone reductase promoter is functional in tobacco, a plant which does not synthesize isoflavonoids. Infection of transgenic alfalfa plants by Phoma causes an increase in -glucuronidase staining, as does elicitation of transgenic alfalfa cell cultures, indicating that this promoter fusion is a good indicator of phytoalexin biosynthesis in alfalfa.Abbreviations CA4H
cinnamic acid 4-hydroxylase
- CHI
chalcone isomerase
- CHOMT
chalcone O-methyltransferase
- CHS
chalcone synthase
- 4CL
4-coumarate:CoA ligase
- COMT
caffeic acid O-methyltransferase
- FGM
malonylated glucoside of formononetin
- GUS
-glucuronidase
- IFOH
isoflavone 2-hydroxylase
- IFR
isoflavone reductase
- IFS
isoflavone synthase
- IOMT
isoflavone 4-O-methyltransferase
- MGM
medicarpin 3-O-glucoside-6-O-malonate
- PAL
L-phenylalanine ammonia-lyase
- PTS
pterocarpan synthase
- VAM
vesicular arbuscular mycorrhizal
- X-gluc
5-bromo-4-chloro-3-indolyl--D-glucuronide 相似文献
11.
12.
A glyceraldehyde-3-phosphate dehydrogenase gene (gpd) promoter (PMagpd) was obtained from Metarhizium acridum and its active region analyzed by 5′-deletion strategy using β-glucuronidase (GUS) as a reporter. Sequence analysis revealed
that typical regulatory elements of PMagpd were included in the 1.7 kb region upstream of the start codon of the Magpd gene. Deletion of the region from −1,691 bp to −1,463 bp, where the gpd box is harbored, did not significantly affect the PMagpd activity. Deletions of the regions upstream of −946 bp and upstream of −684 bp caused a major decrease of GUS activity. Compared
with PgpdA (2.2 kb) in Aspergillus nidulans, PMagpd (1.4 kb) had a shorter sequence and significantly higher activity in M. acridum. This study provides an applicable promoter for over-expression of target genes in M. acridum. 相似文献
13.
A full-length genomic clone of 2,233 bp long containing an anther- and tapetum-specific gene TomA108 was isolated and characterized from tomato. The gene was present in one copy per haploid genome. The isolated clone contained
5′ and 3′ untranslated regions of 810 and 170 nucleotides, respectively and a single intron with highly repetitive sequences.
The cDNA encoded the protein with an apparent mass of 10.6 kDa and a pI (isoelectric point) of 5.3. It was cysteine-rich and
had an N-terminal hydrophobic domain with characteristics of a secretory signal. Amino acid sequence comparisons demonstrated
that the protein was closely related to a family of cereal seed storage proteins and protease inhibitors. The fusion of β-glucuronidase
to the TomA108 promoter demonstrated that the promoter was highly active from early-meiosis to free microspores production in tapetum of
tobacco. This strong and highly specific promoter can be potentially used to generate male sterility for efficient production
of plant hybrids. 相似文献
14.
Molecular Cloning and Characterization of the Myostatin Gene in Croceine Croaker, Pseudosciaena crocea 总被引:1,自引:0,他引:1
Myostatin (MSTN) is a negative regulator of skeletal muscle mass and has a potential application in aquaculture. We reported
the characterization of the myostatin gene and its expression in the croceine croaker, Pseudosciaena crocea. The myostatin gene had three exons encoding 376 amino acids. The cDNA was 1,906 bp long with a 5′-UTR and 3′-UTR of 108 bp
and 667 bp, respectively. A microsatellite sequence, CA30 and CA26 separated by TA, existed in the 3′-UTR. Intron I and II were 343 bp and 758 bp in length, respectively. The deduced amino
acid sequence was highly conserved, and had more than 90% identical to shi drum, gilthead seabream, striped sea-bass, white
perch, and white bass proteins. The myostatin of croceine croaker had a putative amino terminal signal sequence (residues
1–22), a transforming growth factor-beta (TGF-β) propeptide domain (residues 41–256), a RXXR proteolytic processing site (RARR,
residues 264–267, matching the RXXR consensus site), and a TGF-β domain (residues 282–376). There were 13 conserved cysteine
residues in croceine croaker myostatin, nine of which are common to all TGF-β superfamily members. The most conserved region
of vertebrate myostatins is the TGF-β domain, which was the mature bioactive domain of the myostatin protein. The myostatin
gene was expressed not only in the skeletal muscle, but also in the other tissues. 相似文献
15.
Eva Stöger Rosa Maria Benito Moreno Bauke Ylstra Oscar Vicente Erwin Heberle-Bors 《Transgenic research》1992,1(2):71-78
The particle gun, cocultivation withAgrobacterium tumefaciens, and imbibition in DNA solutions were compared as methods to transfer DNA into mature and immature pollen ofNicotiana tabacum. Bombardment of mature pollen with the β-glucuronidase gene cloned behind the pollen-specific PA2 promoter of the chalcone
isomerase gene ofPetunia hybrida resulted in the expression of the β-glucuronidase gene in 0.025% of the pollen grains. Bombardment of younger stages followed
byin vitro maturation also resulted in the formation of mature pollen that expressed β-glucuronidase, although at a lower frequency.
Cocultivation of pollen duringin vitro maturation orin vitro germination withAgrobacterium tumefaciens did not yeild β-glucuronidase-expressing pollen. In these cases, an intron-containing β-glucuronidase gene was used which
effectively prevented β-glucuronidase expression in the bacteria. Imbibition of mature, dry pollen in various DNA solutions
of the same constructs also did not lead to the formation of β-glucuronidase expressing pollen. 相似文献
16.
We have characterized the promoter specificity of theArabidopsis thaliana α1-tubulin (α
1-tub) gene by studying expression patterns of gene fusions between the 2.2 kbp 5′ upstream region of theα
1-tub gene and each of three different reporters: chloramphenical acetyltransferase, β-glucuronidase or the diphtheria toxin chain
A gene. Analysis of transgenic tobacco andArabidopsis plants carrying the transgene showed that the chloramphenicol acetyltransferase and β-glucuronidase activities were not detected
in any vegetative or reproductive organs except mature pollen. Transgenic tobacco plants carrying the diphtheria toxin chain
A gene under the control of theα
1-tub promoter were of normal phenotype but seed fertility was drastically reduced. Furthermore, the transgene could not be transmitted
to the next generation through pollen, supporting the observation that theα
1-tub promoter is active only in pollen. It was observed that the promoter activity was most active in mature pollen and decreased
significantly duringin vitro pollen germination, indicating that the promoter is inactive or subdued in germinating pollen. The promoter activity was
not affected by various plant growth hormones during pollen maturation. 相似文献
17.
18.
19.
Cassettes for seed-specific expression tested in transformed embryogenic cultures of soybean 总被引:4,自引:0,他引:4
Soybean (Glycine max [L.] Merrill) lectin is a seed protein that accumulates in protein bodies of cotyledons during seed development. We have
constructed two expression cassettes containing the 5′ and 3′ regions of the soybean lectin gene connected by aNot I restriction site. One vector also contains the 32 amino acid signal sequence. Using polymerase chain reaction (PCR), the
coding region of the β-glucuronidase (uidA) gene was inserted into theNot I site of each vector. We tested the function of the expression cassettes in transformed embryogenic cultures of soybean.
Development-specific GUS expression was observed in developing somatic embryos transformed with the chimeric lectin promoter-GUS
constructs as determined by histochemical assays. Our data indicate that these cassettes could be used to drive expression
of foreign genes to modify embryo-specific traits of soybean as protein quality or quantity in the seed. 相似文献