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1.
SBgLR (Solanum tuberosum genomic lysine-rich) is a pollen-specific gene cloned from potato (Solanum tuberosum L.). The region from −269 to −9 (The A of translation start site “ATG” as +1) of the SBgLR promoter was identified as critical for gene specific expression in pollen grains. Sequence analysis indicates a palindromic
sequence “TTTCTATTATAATAGAAA” in the −227 to −209 region, in which two pollen-specific motifs TTTCT and AGAAA surround a unique
putative TATA box. Moreover, nine putative pollen-specific motifs are located in the region between the TATA box and ATG.
We placed the −227 to −9 region (reserving the palindrome) and the −222 to −9 region (breaking the palindrome) downstream
of the CaMV35S enhancer, respectively, to construct two fusion promoters. Histochemical assays in transgenic plants demonstrated that the
region from −222 to −9 is necessary and sufficient for pollen-specific expression of the uidA gene. However, the region of −227 to −9 is incapable of driving GUS expression in pollen grains and parts of vegetative tissues.
A series of 5′ deletions from −269 to −9 of SBgLR promoter were constructed. A transient expression assay indicated that the region from the −227 to −9 suppressed gfp gene expression in pollen, and a positive regulatory element was present in the region of −253 to −227. The function of the
palindromic sequence as a repressor inhibiting gene expression in pollen was further confirmed by the mutated promoter, breaking
the palindrome by substituting its 3′-flanking five base pairs, which resumes the reporter gene expression in mature pollen. 相似文献
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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
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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
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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
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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
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Vahid Omidvar Siti Nor Akmar Abdullah Amir Izadfard Chai Ling Ho Maziah Mahmood 《Planta》2010,232(4):925-936
The 1,053-bp promoter of the oil palm metallothionein gene (so-called MSP1) and its 5′ deletions were fused to the GUS reporter
gene, and analysed in transiently transformed oil palm tissues. The full length promoter showed sevenfold higher activity
in the mesocarp than in leaves and 1.5-fold more activity than the CaMV35S promoter in the mesocarp. The 1,053-bp region containing
the 5′ untranslated region (UTR) gave the highest activity in the mesocarp, while the 148-bp region was required for minimal
promoter activity. Two positive regulatory regions were identified at nucleotides (nt) −953 to −619 and −420 to −256 regions.
Fine-tune deletion of the −619 to −420 nt region led to the identification of a 21-bp negative regulatory sequence in the
−598 to −577 nt region, which is involved in mesocarp-specific expression. Gel mobility shift assay revealed a strong interaction
of the leaf nuclear extract with the 21-bp region. An AGTTAGG core-sequence within this region was identified as a novel negative
regulatory element controlling fruit-specificity of the MSP1 promoter. Abscisic acid (ABA) and copper (Cu2+) induced the activity of the promoter and its 5′ deletions more effectively than methyl jasmonate (MeJa) and ethylene. In
the mesocarp, the full length promoter showed stronger inducibility in response to ABA and Cu2+ than its 5′ deletions, while in leaves, the −420 nt fragment was the most inducible by ABA and Cu2+. These results suggest that the MSP1 promoter and its regulatory regions are potentially useful for engineering fruit-specific
and inducible gene expression in oil palm. 相似文献
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Parasites exert a selection pressure on their hosts and are accountable for driving diversity within gene families and immune
gene polymorphisms in a host population. The overwhelming response of regulatory T cells during infectious challenges directs
the host immune system to lose the ability to mount parasite specific T cell responses. The underlying idea of this study
is that regulatory single nucleotide polymorphism (SNPs) can cause significant changes in gene expression in functional immune
genes. We identified and investigated regulatory SNPs in the promoter region of the FOXP3 gene in a group of Gabonese individuals exposed to a variety of parasitic infections. We identified two novel and one promoter
variants in 40 individual subjects. We further validated these promoter variants for their allelic gene expression using transient
transfection assays. Two promoter variants, −794 (C/G) and the other variant −734/−540 (C/T) revealed a significant higher
expression of the reporter gene at basal level in comparison to the major allele. The identification of SNPs that modify function
in the promoter regions could provide a basis for studying parasite susceptibility in association studies. 相似文献
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Dipnarayan Saha Vajinder Kumar Shripad Ramachandra Bhat Ramamurthy Srinivasan 《Plant Molecular Biology Reporter》2011,29(2):265-277
Isolation and characterization of promoters are important in understanding gene regulation and genetic engineering of crop
plants. Earlier, a pentatricopeptide repeat protein (PPR) encoding gene (At2g39230), designated as Lateral Organ Junction (LOJ) gene, was identified through T-DNA promoter trapping in Arabidopsis thaliana. The upstream sequence of the LOJ gene conferred on the reporter gene a novel LOJ-specific expression. The present study was aimed at identifying and characterizing
the cis-regulatory motifs responsible for tissue-specific expression in the −673 and +90 bases upstream of the LOJ gene recognized as LOJ promoter. In silico analysis of the LOJ promoter revealed the presence of a few relevant regulatory motifs and a unique feature like AT-rich inverted repeat. Deletion
analysis of the LOJ promoter confirmed the presence of an enhancer-like element in the distal region (−673/−214), which stimulates a minimal
promoter-like sequence in the −424/−214 region in a position and orientation autonomous manner. The −136/+90 region of the
LOJ promoter was efficient in driving reporter gene expression in tissues like developing anthers and seeds of Arabidopsis. A positive regulation for the seed- and anther-specific expression module was contemplated within the 5′ untranslated region
of the LOJ gene. However, this function was repressed in the native context by the lateral organ junction-specific expression. The present
study has led to the identification of a novel lateral organ junction-specific element and an enhancer sequence in Arabidopsis with potential applications in plant genetic engineering. 相似文献
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K. E. Orishchenko E. A. Elisaphenko A. E. Kel S. M. Zakian 《Russian Journal of Genetics》2009,45(10):1182-1191