scholarly journals Isolation, Cloning and Characterization of a Constitutive Plant from Potato Aquaporin Gene

2020 ◽  
Vol 63 (3) ◽  
pp. 169-178
Author(s):  
Hira Mubeen ◽  
Rubab Zahra Naqvi ◽  
Ammara Masood ◽  
Mushtaq A. Saleem ◽  
Aftab Bashir ◽  
...  
Keyword(s):  
Genomic Sequence ◽  
Crop Improvement ◽  
Regulatory Elements ◽  
Gus Expression ◽  
Cis Acting ◽  
Transcription Start Sites ◽  
Expression Studies ◽  
Aquaporin Gene ◽  
Highly Expressed Genes

Plasma membrane intrinsic proteins (PIP1) are the most common integral membrane proteins belong to a larger family of intrinsic aquaporin proteins. They are member of aquaporin gene family and have gained importance as highly expressed genes in plants. In this study, the promoter of aquaporin PIP1 gene was identified, analyzed and retrieved from high throughput genomic sequence (HTGS) database. The cis-acting regulatory elements, transcription start sites and transcription factor binding sites of selected promoter were identified through different bio-informatics tools. Many light responsive, phytohormone, stress and defense related cis-regulatory elements were detected in PIP1 promoter region indicating its role as a constitutive promoter. The PIP1 promoter was isolated from Solanum tuberosum. It was initially cloned in TA vector (pTZ57R/T) and later transferred to plant expression binary vectors, pGR1 and pGA482 for transient and stable expression studies in tobacco. The GUS expression results of PIP1 promoter in different tobacco tissues showed its functional importance in regulating gene expression in a constitutive manner. Further, it was concluded that the PIP1 aquaporin promoter is constitutively expressed with a strength equivalent to CaMV 2x35S promoter. These findings indicated the significance of isolated promoter for genetic engineering of plants for crop improvement.  

Cloning and characterization of the light-inducible Gacab promoter from Gossypium arboreum

2005 ◽  
Vol 2 (1) ◽  
pp. 17-22 ◽  
Author(s):  
Li Wei-Min ◽  
Wang Zhi-Xing ◽  
Pei Xin-Wu ◽  
Jia Shi-Rong
Keyword(s):  
Promoter Sequence ◽  
Inducible Promoter ◽  
Regulatory Elements ◽  
Gus Expression ◽  
Full Length ◽  
Expression Vectors ◽  
35S Promoter ◽  
Gossypium Arboreum ◽  
Gus Histochemical Assay

AbstractA 1009 bp promoter sequence of cab gene, which encodes chlorophyll a/b binding protein belonging to a class of light-inducible proteins, was cloned from Gossypium arboreum. Sequence analysis showed that it had no obvious homology with previously published cab promoters. The full-length Gacab promoter and 5′ deletions with length of 197, 504 and 779 bp were fused with gus (uid A) gene, respectively, and plant expression vectors were used for transformation of Nicotiana tabacum cv. NC89. β-Glucuronidase (GUS) histochemical assay of transgenic tobacco plants showed that GUS was expressed specifically in leaves and young green tissues. GUS was not detected in the leaves of transgenic plants grown in the dark for 6 days. However, it was highly expressed in the leaves of these plants after induction with light for another 6 days, demonstrating that the full-length Gacab promoter is a light-inducible promoter. Transient GUS expression in rice calli indicated that the expression level of Gacab504::gus was the highest and stronger than that of the CaMV 35S promoter, while expression was reduced for Gacab197::gus, Gacab779::gus and Gacab1009::gus constructs. This suggests that −197 bp to −1 bp is a basic promoter of Gacab, some positive regulatory elements may exist in −504 bp to −197 bp, and the fragment −1009 bp to −504 bp may contain negative elements.

scholarly journals Hepatic transcription of the acute-phase alpha 1-inhibitor III gene is controlled by a novel combination of cis-acting regulatory elements.

1990 ◽  
Vol 10 (7) ◽  
pp. 3483-3491 ◽  
Author(s):  
L J Abraham ◽  
A D Bradshaw ◽  
B R Shiels ◽  
W Northemann ◽  
G Hudson ◽  
...  
Keyword(s):  
Binding Sites ◽  
Transcriptional Start Site ◽  
Regulatory Elements ◽  
Nuclear Factor ◽  
Base Pairs ◽  
Cis Acting ◽  
Expression Studies ◽  
Positive Regulators ◽  
Flanking Region ◽  
Related Proteins

mRNA coding for the abundant broad-range plasma proteinase inhibitor alpha 1-inhibitor III (alpha 1I3) was detected only in rat liver, while mRNA for the related proteins alpha 1-macroglobulin and alpha 2-macroglobulin was also found in a variety of nonhepatic tissues. cis-Acting control elements necessary for the hepatic transcription of alpha 1I3 were mapped by transfection and expression studies of control-region constructs in cultured hepatic and nonhepatic cells. The promoter-proximal 5'-flanking region contained four control elements, I to IV, located between -109 and -196 base pairs upstream of the transcriptional start site relevant for the hepatic transcription of this gene. Elements II and III were essential, and I and IV exerted strong modulatory effects. Elements I to III acted as positive regulators, and IV acted as a negative element. Element II contained the sequence TGGCA and is probably a binding site for a nuclear factor related to the known transcription factor NF1. The other three elements did not resemble consensus binding sites for known transcription factors that are involved in the hepatocyte-specific transcription of other well-characterized plasma protein genes, such as the prototype factor HNF-1. Thus, the alpha 1I3 gene achieves its highly hepatocyte-specific transcription through a novel combination of cis-acting control elements and trans-acting factors.

SlPMEI, a pollen-specific gene in tomato

2014 ◽  
Vol 94 (1) ◽  
pp. 73-83 ◽  
Author(s):  
Woong Bom Kim ◽  
Chan Ju Lim ◽  
Hyun A. Jang ◽  
So Young Yi ◽  
Sang-Keun Oh ◽  
...  
Keyword(s):  
Specific Activity ◽  
Pectin Methylesterase ◽  
Gus Expression ◽  
Specific Gene ◽  
Specific Expression ◽  
Cis Acting ◽  
Solanum Lycopersicum L ◽  
Flower Organs ◽  
Main Components

Kim, W. B., Lim, C. J., Jang, H. A., Yi, S. Y., Oh, S.-K., Lee, H. Y., Kim, H. A., Park, Y.-I. and Kwon, S.-Y. 2014. SlPMEI, a pollen-specific gene in tomato. Can. J. Plant Sci. 94: 73–83. Pectin is one of the main components of plant cell walls, and its biosynthesis is controlled by pectin methylesterase (PME). Pectin methylesterase inhibitors (PMEIs) are key regulators of PME. We report here the cloning and characterization of a novel Solanum lycopersicum L. PMEI gene, SlPMEI. RT-PCR studies of leaf, seed, fruit, flower, and flower organs confirmed that SlPMEI is expressed specifically in pollen. Promoter analysis of SlPMEI revealed pollen-specific cis-acting elements (pollen lat52 and g10). In addition, SlPMEI is expressed independently of abiotic stress, pathogen exposure, and growth stage in tomato, and a histochemical analysis of promoter activity revealed pollen-specific expression in both Arabidopsis and tomato. Under the microscope, we observed pollen-specific GUS expression in the stamen of transgenic tomato plant. These results indicate that the promoter of SlPMEI has strong pollen-specific activity, and could therefore be useful for development of industrially and agronomically important transgenic plants.

Hepatic transcription of the acute-phase alpha 1-inhibitor III gene is controlled by a novel combination of cis-acting regulatory elements

1990 ◽  
Vol 10 (7) ◽  
pp. 3483-3491
Author(s):  
L J Abraham ◽  
A D Bradshaw ◽  
B R Shiels ◽  
W Northemann ◽  
G Hudson ◽  
...  
Keyword(s):  
Binding Sites ◽  
Transcriptional Start Site ◽  
Regulatory Elements ◽  
Nuclear Factor ◽  
Base Pairs ◽  
Cis Acting ◽  
Expression Studies ◽  
Positive Regulators ◽  
Flanking Region ◽  
Related Proteins

mRNA coding for the abundant broad-range plasma proteinase inhibitor alpha 1-inhibitor III (alpha 1I3) was detected only in rat liver, while mRNA for the related proteins alpha 1-macroglobulin and alpha 2-macroglobulin was also found in a variety of nonhepatic tissues. cis-Acting control elements necessary for the hepatic transcription of alpha 1I3 were mapped by transfection and expression studies of control-region constructs in cultured hepatic and nonhepatic cells. The promoter-proximal 5'-flanking region contained four control elements, I to IV, located between -109 and -196 base pairs upstream of the transcriptional start site relevant for the hepatic transcription of this gene. Elements II and III were essential, and I and IV exerted strong modulatory effects. Elements I to III acted as positive regulators, and IV acted as a negative element. Element II contained the sequence TGGCA and is probably a binding site for a nuclear factor related to the known transcription factor NF1. The other three elements did not resemble consensus binding sites for known transcription factors that are involved in the hepatocyte-specific transcription of other well-characterized plasma protein genes, such as the prototype factor HNF-1. Thus, the alpha 1I3 gene achieves its highly hepatocyte-specific transcription through a novel combination of cis-acting control elements and trans-acting factors.

Characterization of cis-acting elements regulating transcription from the promoter of a constitutively active rice actin gene

1992 ◽  
Vol 12 (8) ◽  
pp. 3399-3406
Author(s):  
Y Wang ◽  
W Zhang ◽  
J Cao ◽  
D McElroy ◽  
R Wu
Keyword(s):  
Transient Expression ◽  
Plant Cell ◽  
Transcription Initiation ◽  
Regulatory Elements ◽  
Initiation Site ◽  
Gus Expression ◽  
Negative Regulator ◽  
Cis Acting ◽  
Highly Active ◽  
Constitutively Active

The promoter of the constitutively expressed rice (Oryza sativa) actin 1 gene (Act1) is highly active in transformed rice plants (W. Zhang, D. McElroy, and R. Wu, Plant Cell 3:1150-1160, 1991). A region 834 bp upstream of the Act1 transcription initiation site contains all the regulatory elements necessary for maximal gene expression in transformed rice protoplasts (D. McElroy, W. Zhang, J. Cao, and R. Wu, Plant Cell 2:163-171, 1990). We have constructed a series of Act1 promoter deletions fused to a bacterial beta-glucuronidase reporter sequence (Gus). Transient expression assays in transformed rice protoplasts, as well as transformed maize cells and tissues, identified two distinct cis-acting regulatory elements in the Act1 promoter. A 38-bp poly(dA-dT) region was found to be a positive regulator of Act1 promoter activity. Deletion of the poly(dA-dT) element lowered Gus expression by at least threefold compared with expression produced by the full-length Act1 promoter. By gel retardation and footprinting, we identified a ubiquitous rice protein which specifically recognizes this poly(dA-dT) element in the constitutively active Act1 promoter. A CCCAA pentamer repeat-containing region was found to be a negative regulator of the Act1 promoter in transformed rice protoplasts. Transient expression assays in different maize cells and tissues with use of the Act1 deletion constructs suggested that the CCCAA pentamer repeat region functions in a complex tissue-specific manner. A CCCAA-binding protein was detected only in root extracts.

scholarly journals Identification and Characterization of the Glutathione Peroxidase (GPX) Gene Family in Watermelon and Its Expression under Various Abiotic Stresses

Agronomy ◽  
2018 ◽  
Vol 8 (10) ◽  
pp. 206 ◽  
Author(s):  
Yong Zhou ◽  
Jingwen Li ◽  
Junhong Wang ◽  
Wenting Yang ◽  
Youxin Yang
Keyword(s):  
Abiotic Stress ◽  
Glutathione Peroxidase ◽  
Regulatory Elements ◽  
Promoter Regions ◽  
Cis Acting ◽  
Genome Wide ◽  
A Genome ◽  
Identification And Characterization ◽  
Lower Expression

Plant glutathione peroxidase (GPX) is an important antioxidant enzyme to maintain H2O2 homeostasis and regulate plant response to abiotic stress. In this paper, we present the first report of a genome-wide identification of GPX genes in watermelon. A total of six genes (ClGPX1–ClGPX6) were identified, which were unevenly located on four chromosomes of the watermelon genome. Based on phylogenetic analysis, the GPX genes of Arabidopsis, rice, cucumber, and sorghum were classified into four groups. Through analyzing the promoter regions of ClGPX genes, many development-, stress-, and hormone-responsive cis-acting regulatory elements were also identified. Expression pattern analysis by qRT-PCR indicated that all ClGPX genes were actively expressed in flowers and fruits, and exhibited relatively lower expression in other tissues, particularly roots and stems. In addition, the expression of ClGPX genes was significantly induced by salt, drought, and cold stresses, as well as abscisic acid (ABA) treatment at different time points, suggesting that they may be involved in response to abiotic stress and ABA. Taken together, our findings demonstrated that ClGPX genes might function in watermelon development, especially in flower and fruit tissue, as well as in response to abiotic stress and hormones.

scholarly journals Characterization of cis-acting elements regulating transcription from the promoter of a constitutively active rice actin gene.

1992 ◽  
Vol 12 (8) ◽  
pp. 3399-3406 ◽  
Author(s):  
Y Wang ◽  
W Zhang ◽  
J Cao ◽  
D McElroy ◽  
R Wu
Keyword(s):  
Transient Expression ◽  
Plant Cell ◽  
Transcription Initiation ◽  
Regulatory Elements ◽  
Initiation Site ◽  
Gus Expression ◽  
Negative Regulator ◽  
Cis Acting ◽  
Highly Active ◽  
Constitutively Active

The promoter of the constitutively expressed rice (Oryza sativa) actin 1 gene (Act1) is highly active in transformed rice plants (W. Zhang, D. McElroy, and R. Wu, Plant Cell 3:1150-1160, 1991). A region 834 bp upstream of the Act1 transcription initiation site contains all the regulatory elements necessary for maximal gene expression in transformed rice protoplasts (D. McElroy, W. Zhang, J. Cao, and R. Wu, Plant Cell 2:163-171, 1990). We have constructed a series of Act1 promoter deletions fused to a bacterial beta-glucuronidase reporter sequence (Gus). Transient expression assays in transformed rice protoplasts, as well as transformed maize cells and tissues, identified two distinct cis-acting regulatory elements in the Act1 promoter. A 38-bp poly(dA-dT) region was found to be a positive regulator of Act1 promoter activity. Deletion of the poly(dA-dT) element lowered Gus expression by at least threefold compared with expression produced by the full-length Act1 promoter. By gel retardation and footprinting, we identified a ubiquitous rice protein which specifically recognizes this poly(dA-dT) element in the constitutively active Act1 promoter. A CCCAA pentamer repeat-containing region was found to be a negative regulator of the Act1 promoter in transformed rice protoplasts. Transient expression assays in different maize cells and tissues with use of the Act1 deletion constructs suggested that the CCCAA pentamer repeat region functions in a complex tissue-specific manner. A CCCAA-binding protein was detected only in root extracts.

scholarly journals Molecular cloning and characterization of a lipoxygenase gene (CsLOX1) from cucumber

2020 ◽  
Vol 48 (4) ◽  
pp. 1832-1844
Author(s):  
Yong ZHOU ◽  
Mingyuan XU ◽  
Wei LAI ◽  
Lianghai CHEN ◽  
Yingui YANG ◽  
...  
Keyword(s):  
Defense Responses ◽  
Regulatory Elements ◽  
Heme Iron ◽  
Male Sterile ◽  
Multiple Sequence ◽  
Conserved Residues ◽  
Cis Acting ◽  
Non Heme Iron ◽  
Lipoxygenase Gene

Lipoxygenases (LOXs) are non-heme iron enzymes that play crucial roles in many developmental processes during plant life, and defense responses against biotic and abiotic stresses. In this study, a lipoxygenase gene (CsLOX1) was cloned and characterized from cucumber (Cucumis sativus). The coding sequence (CDS) of CsLOX1 was 741 bp, and encoded an 878 amino-acid residue protein, which was predicted to be located in the cytoplasm. CsLOX1 contained the conserved LH2/PLAT and lipoxygenase domains, as well as the representative 38 amino acids motif [His-(X)4-His-(X)4-His-(X)17-His-(X)8-His]. Multiple sequence alignment and phylogenetic analysis indicated that CsLOX1 was closely related to other dicot 9-LOXs and posesess the essential conserved residues involved in the binding of the iron atom. Promoter analysis suggested that several development-, stress-, and hormone-related cis-acting regulatory elements were present in the promoter region of CsLOX1. The function of CsLOX1 was assessed by overexpression it in Arabidopsis, and the transgenic plants were male sterile and displayed obviously increased floral shoots. These results provide some clues for revealing the biological roles of CsLOX1 in cucumber.

scholarly journals Abiotic stress responsive cis-regulatory elements (CREs) in rice (Oryza sativa L.) and other plants

2019 ◽  
Author(s):  
Sangram Lenka ◽  
Kailash C Bansal
Keyword(s):  
Gene Expression ◽  
Abiotic Stress ◽  
Large Scale ◽  
Sequence Data ◽  
Oryza Sativa L ◽  
Crop Improvement ◽  
Regulatory Elements ◽  
Plant Genome ◽  
Agricultural Crop ◽  
Cis Acting

Capability of crop plants to adjust to the adverse environmental conditions in a spatiotemporalfashion is critical for their survival and maintaining agricultural productivity.Genetic engineering efforts for improving tolerance to diverse abiotic stresses in crop plantsusing well characterised stress-inducible promoter elements have proven to be advantageous.Combinatorial interactions of cis-acting DNA elements in the promoters with trans-actingprotein factors are key processes governing spatio-temporal gene expression. It is becomingincreasingly evident that targeted modification of molecular genetic network is feasible, forexploiting the potential of specific abiotic stress responsive element and its correspondingmaster regulatory genes via plant genetic engineering.The importance of inducible promotersin agricultural crop improvement is enormous; hence it is very crucial to characteriseinducible promoters from plant genome sequence data bases on a large scale. We will brieflydiscuss here abiotic stress responsive cis-acting elements and their role in abiotic stressregulated gene expression.

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