共查询到20条相似文献,搜索用时 437 毫秒
1.
Rastogi Rajeev Bate Nicholas J. Sivasankar Sobhana Rothstein Steven J. 《Plant molecular biology》1997,34(3):465-476
Nitrite reductase (NiR) is the second enzyme in the nitrate assimilatory pathway reducing nitrite to ammonium. The expression of the NiR gene is induced upon the addition of nitrate. In an earlier study, a 130 bp upstream region of the spinach NiR gene promoter, located between –330 to –200, was shown to be necessary for nitrate induction of -glucuronidase (GUS) expression in tissue-specific manner in transgenic tobacco plant [28]. To further delineate the cis-acting elements involved in nitrate regulation of NiR gene expression, transgenic tobacco plants were generated with 5 deletions in the–330 to –200 region of the spinach NiR gene promoter fused to the GUS gene. Plants with the NiR promoter deleted to –230 showed a considerable increase in GUS activity in the presence of nitrate, indicating that the 30 bp region between –230 to –200 is crucial for nitrate-regulated expression of NiR. In vivo DMS footprinting of the –300 to –130 region of the NiR promoter in leaf tissues from two independent transgenic lines revealed several nitrate-inducible footprints. Footprinting within the –230 to –181 region revealed factor binding to two adjacent GATA elements separated by 24 bp. This arrangement of GATA elements is analogous to cis-regulatory sequences found in the promoters of nitrate-inducible genes of Neurospora crassa, regulated by the NIT2 Zn-finger protein. The –240 to –110 fragment of the NiR promoter, which contains two NIT2 consensus core elements, bound in vitro to a fusion protein comprising the zinc finger domain of the N. crassa NIT2 protein. The data presented here show that nitrate-inducible expression of the NiR gene is mediated by nitrate-specific binding of trans-acting factors to sequences preserved between fungi and higher plants. 相似文献
2.
Jocelyne Kronenberger Andrée Lepingle Michel Caboche Hervé Vaucheret 《Molecular & general genetics : MGG》1993,236(2-3):203-208
Summary Three tobacco nitrite reductase (NiR) cDNA clones were isolated using spinach NiR cDNA as a probe. Sequence analysis and Southern blot hybridization revealed four genes in tobacco. Two of these genes presumably derived from the ancestral species Nicotiana tomentosiformis, the other two from the ancestor N. sylvestris. Northern blot analysis showed that one gene from each ancestral genome was expressed predominantly in leaves, whilst RNA from the other was detected mostly in roots. The accumulation of both leaf and root NiR mRNAs was induced by nitrate and repressed by nitrate- or ammonium-derived metabolites. In addition, the expression of the root NiR gene was detectable in leaves of a tobacco nitrate reductase (NR)-deficient mutant. Thus, the regulation of expression of tobacco NiR genes is comparable to the regulation of expression of barley NR genes. 相似文献
3.
4.
A 330 bp region of the spinach nitrite reductase gene promoter directs nitrate-inducible tissue-specific expression in transgenic tobacco 总被引:4,自引:0,他引:4
Rajeev Rastogi Eduard Back Alois Schneiderbauer Caroline G. Bowsher Barbara Moffatt Steven J. Rothstein 《The Plant journal : for cell and molecular biology》1993,4(2):317-326
5.
Zhao-Jun Wei Miao Yu Shun-Ming Tang Yong-Zhu Yi Gui-Yun Hong Shao-Tong Jiang 《Molecular biology reports》2011,38(2):1121-1127
Prothoracicotropic hormone (PTTH) is one of key players in regulation of insect growth, molting, metamorphosis, diapause,
and is expressed specifically in the two pairs of lateral PTTH-producing neurosecretory cells in the brain. Analysis of cis-regulatory elements of the PTTH promoter might elucidate the regulatory mechanism controlling PTTH expression. In this study,
the PTTH gene promoter of Bombyx mori (Bom-PTTH) was cloned and sequenced. The cis-regulatory elements in Bom-PTTH gene promoter were predicted using Matinspector software, including myocyte-specific enhancer
factor 2, pre-B-cell leukemia homeobox 1, TATA box, etc. Transient transfection assays using a series of fragments linked
to the luciferase reporter gene indicated that the fragment spanning −110 to +33 bp of the Bom-PTTH promoter showed high ability
to support reporter gene expression, but the region of +34 to +192 bp and −512 to −111 bp repressed the promoter activity
in the BmN and Bm5 cell lines. Electrophoretic mobility shift assays demonstrated that the nuclear protein could specifically
bind to the region spanning −124 to −6 bp of the Bom-PTTH promoter. Furthermore, we observed that the nuclear protein could
specifically bind to the −59 to −30 bp region of the Bom-PTTH promoter. A classical TATA box, TATATAA, localized at positions
−47 to −41 bp, which is a potential site for interaction with TATA box binding protein (TBP). Mutation of this TATA box resulted
in no distinct binding band. Taken together, TATA box was involved in regulation of PTTH gene expression in B. mori. 相似文献
6.
7.
8.
9.
10.
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
11.
We isolated and characterized a pollen-preferential gene, BAN102, from Chinese cabbage and analyzed the activity of its promoter. There were three or four copies of the BAN102 gene in the Chinese cabbage genome that specifically expressed in pollen and pollen tube. There were 2137 bp of BAN102 genomic clone comprising 186 bp of protein coding region, and 1178 bp of 5′ and 773 bp of 3′ non-coding regions. TATA box
were located at 1071 nt of the promoter region while the polyadenylation signal and polyadenylation site were at 1470 and
1486 nt of the 3′ non-coding region. BLAST search of BAN102 sequence showed that coding region of BAN102 gene was the greatest percent similarity with arabinogalactan protein (AGP23) gene from Arabidopsis thaliana. Promoter analysis using GUS gene as a reporter showed that the pollen-specificity of BAN102 resided within the −112 to −44 bp of proximal promoter from the transient expression in tobacco and Chinese cabbage plants. 相似文献
12.
Faustino Merchán Rafael Prieto Karen L. Kindle María J. Llama Juan L. Serra Emilio Fernández 《Plant molecular biology》1995,27(5):1037-1042
The nitrite reductase (NiR) gene (nirA) has been isolated and sequenced from the filamentous, thermophilic non-N2-fixing cyanobacterium Phormidium laminosum. Putative promoter-like and Shine-Dalgarno sequences appear at the 5 end of the 1533 bp long nir-coding region. The deduced amino acid sequence of NiR from P. laminosum corresponds to a 56 kDa polypeptide, a size identical to the molecular mass previously determined for the pure enzyme, and shows a high identity with amino acid sequences from ferredoxin-dependent NiR. This cyanobacterial NiR gene has been efficiently expressed in Escherichia coli DH5 from the E. coli lac promoter and probably from the P. laminosum NiR promoter.Abbreviations IPTG
isopropyl--D-thiogalactopyranoside
- NiR
nitrite reductase
- NR
nitrate reductase
- NT
nitrate transport
- SiR
sulfite reductase 相似文献
13.
14.
15.
SBgLR (Solanum tuberosum genomic lysine-rich) gene was isolated from a potato genomic library using SB401 (S.
berthaultii 401) cDNA as probe. RT-PCR analysis of SBgLR gene expression profile and microscopic analysis of green fluorescent protein (GFP) expression in tobacco plants transformed
with SBgLR promoter-GFP reporters indicate that SBgLR is a pollen-specific gene. A series of 5′deletions of SBgLR promoter were fused to the β-glucuronidase (GUS) gene and stably introduced into tobacco plants. Histochemical and quantitative assays of GUS expression in transgenic
plants allowed us to localize an enhancer of SBgLR promoter to the region −345 to −269 relative to the translation start site. This 76 bp (−345 to −269) fragment enhanced GUS
expression in leaves, stems and roots when fused to −90/+6 CaMV 35S minimal promoter. Deletion analysis showed that a cis-element, which can repress gene expression in root hairs, was located in the region −345 to −311. Further study indicated
that the −269 to −9 region was sufficient to confer pollen-specific expression of GFP when fused to CaMV 35S enhancer.
Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.
Authors Zhihong Lang and Peng Zhou contributed equally to this work. 相似文献
16.
Xin-Wen Hu Si-Xin Liu Jian-Chun Guo Ji-Tao Li Rui-Jun Duan Shao-Ping Fu 《Functional & integrative genomics》2009,9(3):351-361
Mabinlin II is one of the major sweet proteins stored in the seeds of Capparis masaikai Lévl. Its promoter region (779 bp) located 5′ upstream of the mabinlin II gene has been isolated and named as MBL-779 (GenBank
accession number, EU014073). This promoter contains two typical TATA box regions and a series of motifs related to seed-specific
promoters, such as ACGT motifs, RY motif, napin motif, and G box. The MBL-779 promoter drove GUS gene to transiently express in the embryos of bean, maize, and rice seeds or to constantly express in the embryos and anthers
of the transgenic Arabidopsis. The MBL-779 promoter regulated gene expression from approximately the 12th day and peaked on approximately the 16th day
after flowering in Arabidopsis. The −300-bp promoter region is a minimal sequence required to functionally regulate gene expression. The CAATs at −325 to
−322 bp and −419 to −416 bp and the region at −485 to −770 bp play a role in the quantitative regulation of gene expression.
The RY motif, CATGAC, at −117 to −112 bp and the ACGT within the G box (CACGTG) at −126 to −123 bp positively regulate gene
expression.
X.-W. Hu and S.-X. Liu have the same contribution as first author. 相似文献
17.
The interaction of sulfate assimilation with nitrate assimilation inBrassica juncea roots was analyzed by monitoring the regulation of ATP sulfurylase (AS), adenosine-5’-phosphosulfate reductase (AR), sulfite
reductase (SiR), and nitrite reductase (NiR). Depending on the status of sulfur and nitrogen nutrition, AS and AR activities
and mRNA levels were increased by sulfate starvation but decreased by nitrate starvation. The activation of AS and AR by sulfate
starvation was inhibited by sulfate/nitrate starvation. However, the rise in steady-state mRNA levels for AS and AR by sulfate
starvation was not affected by sulfate/nitrate starvation. SiR gene expression was slightly activated by both sulfate starvation
and sulfate/nitrate starvation, but was decreased by nitrate starvation. Although NiR gene expression was little affected
by sulfate starvation, it was diminished significantly by either nitrate or nitrate/sulfate starvation. Cysteine (Cys) also
decreased AS and AR activities and mRNA levels even when plants were simultaneously starved for sulfate; in contrast, both
SiR and NiR gene expressions were only slightly, if at all, affected under the same conditions. This supports our conclusion
that Cys, the end-product of sulfate assimilation, is the key regulatory signal. Moreover, SiR and NiR apparently are not
the linking step in the co-regulation of sulfate and nitrate assimilation in plants. 相似文献
18.
P. B. F. Ouwerkerk T. O. Trimborn F. Hilliou J. Memelink 《Molecular & general genetics : MGG》1999,261(4-5):610-622
Plant secondary metabolites of the terpenoid indole alkaloid (TIA) class comprise several compounds with pharmaceutical applications.
A key step in the TIA biosynthetic pathway is catalysed by the enzyme tryptophan decarboxylase (TDC), which channels the primary
metabolite tryptophan into TIA metabolism. In Catharanthus roseus (Madagascar periwinkle), the Tdc gene is expressed throughout plant development. Moreover, Tdc gene expression is induced by external stress signals, such as fungal elicitor and UV light. In a previous study of Tdc promoter architecture in transgenic tobacco it was shown that the −538 to −112 region is a quantitative determinant for the
expression level in different plant organs. Within this sequence one particular region (−160 to −99) was identified as the
major contributor to basal expression and another region (−99 to −37) was shown to be required for induction by fungal elicitor.
Here, the in vitro binding of nuclear factors to the −572 to −37 region is described. In extracts from tobacco and C. roseus, two binding activities were detected that could be identified as the previously described nuclear factors GT-1 and 3AF1,
based on their mobility and binding characteristics. Both factors appeared to interact with multiple regions in the Tdc promoter. Mutagenesis of GT-1 binding sites in the Tdc promoter did not affect the basal or elicitor-induced expression levels. However, induction of the Tdc promoter constructs by UV light was significantly lower, thereby demonstrating a functional role for GT-1 in the induction
of Tdc expression by UV light.
Received: 2 February 1998 / Accepted: 5 March 1999 相似文献
19.
We previously cloned and analyzed the 1,893-bp promoter region (−1,915 to −23) of the tomato (Lycopersicon esculentum) Lehsp23.8 gene, whose expression is induced by treatment with high or low temperatures, heavy metal, or abscisic acid (ABA). In our
present work, we examined how this expression is regulated. A comprehensive quantitative promoter deletion and base-substitution
analysis was conducted under various environmental conditions. The proximal region (−565 to −23 bp) of the Lehsp23.8 promoter harbors cis-regulatory elements that conferred high levels of heat-induced expression in transgenic tobacco. Mutation of the five proximal
HSEs (HSE1 to 5) of that promoter led to an absence of heat inducibility. The AT-rich regions between −255 bp and −565 bp
(AT-rich1 to 4) in the promoter might serve as enhancers for such heat-induced expression. Deletion and HSE mutation analysis
indicated that other cis-acting elements also function in response to low temperature, heavy metal, and ABA and that HSE1 to 5 act at least as cis-acting elements in multiple-stress responses of Lehsp23.8. These results reveal that those five proximal HSEs and AT-rich regions function interdependently in the expression of Lehsp23.8 in response to non-heat stresses. Furthermore, the putative elements CRT/DRE, AP-1, and ABRE in that promoter are not required
for multiple-stress induction. 相似文献
20.
Earlier, a pollen-specific Oryza sativa indica pollen allergen gene (OSIPA), coding for expansins/pollen allergens, was isolated from rice, and its promoter—upon expression in tobacco and Arabidopsis—was found active during the late stages of pollen development. In this investigation, to analyze the effects of different
putative regulatory motifs of OSIPA promoter, a series of 5′ deletions were fused to β-glucuronidase gene (GUS) which were stably introduced into rice and Arabidopsis. Histochemical GUS analysis of the transgenic plants revealed that a 1631 bp promoter fragment mediates maximum GUS expression
at different stages of anther/pollen development. Promoter deletions to −1272, −966, −617, and −199 bp did not change the
expression profile of the pollen specificity. However, the activity of promoter was reduced as the length of promoter decreased.
The region between −1567 and −199 bp was found adequate to confer pollen-specific expression in both rice and Arabidopsis systems. An approximate 4-fold increase in the GUS activity was observed in the pollen of rice when compared to that of Arabidopsis. As such, the OSIPA promoter seems promising for generation of stable male-sterile lines required for the production of hybrids in rice and other
crop plants. 相似文献