scholarly journals Effect of nuclear matrix attachment regions on transgene expression in tobacco plants.

2001 ◽  
Vol 48 (3) ◽  
pp. 637-646 ◽  
Author(s):  
W Nowak ◽  
M Gawłowska ◽  
A Jarmołowski ◽  
J Augustyniak
Keyword(s):  
Transgenic Plants ◽  
Reporter Gene ◽  
Transgene Expression ◽  
Binary Vector ◽  
Special Property ◽  
Matrix Attachment Regions ◽  
Plant Populations ◽  
Tobacco Plants ◽  
Gus Reporter ◽  
Loop Domains

Matrix attachment regions (MARs) are thought to participate in the organization and segregation of independent chromosomal loop domains. Although there are several reports on the action of natural MARs in the context of heterologous genes in transgenic plants, in our study we tested a synthetic MAR (sMAR) with the special property of unpairing when under superhelical strain, for its effect on reporter gene expression in tobacco plants. The synthetic MAR was a multimer of a short sequence from the MAR 3' end of the immunoglobulin heavy chain (IgH) enhancer. This sMAR sequence was used to flank the beta-glucuronidase (GUS) reporter gene within the T-DNA of the binary vector pBI121. Vectors with or without the sMARs were then used to transform tobacco plants by Agrobacterium tumefaciens. Transgenic plants containing the sMAR sequences flanking the GUS gene exhibited higher levels of transgene expression compared with transgenic plants which lacked the sMARs. This effect was observed independently of the position of the sMAR at the 5' side of the reporter gene. However, variation of the detected transgene expression was significant in all transformed plant populations, irrespective of the construct used.

scholarly journals CHARACTERISATION OF HORDEUM VULGARE CELLULOSE SYNTHASE-LIKE F6 PROMOTER VIA TRANSGENE EXPRESSION IN RICE

2021 ◽  
Vol 40 (2) ◽  
pp. 61-86
Author(s):  
Azreena Jamahari ◽  
Wong Ling-Chie ◽  
Fan Xioalai ◽  
Liu Qiaoquan ◽  
Leong Sui Sien ◽  
...  
Keyword(s):  
Reporter Gene ◽  
Transgene Expression ◽  
Blood Cholesterol ◽  
Endosperm Tissue ◽  
Beta Glucan ◽  
Glucan Synthase ◽  
Rice Endosperm ◽  
Transgenic Expression ◽  
Gus Reporter Gene ◽  
Gus Reporter

Beta-glucan in cereal crops is known as a functional food, which can reduce cardiovascular diseases by lowering blood cholesterol levels. However, beta-glucan content is relatively low in rice grains, despite being relatively abundant in barley and oat grains. Taking advantage of rice as the staple food for Asians, increasing beta-glucan content in rice for their consumption may help to reduce cardiovascular-related diseases among them. Previous attempts in increasing beta-glucan content in rice via transgene expression of beta-glucan synthase genes from barley into rice were unsuccessful due to the use of non-tissue specific as well as constitutively expressing promoter. The current transgenic expression study was performed to characterise the promoter of beta-glucan synthase gene in barley using beta-glucuronidase (GUS) reporter gene. Two fragments of HvCslF6 promoter (2771 bp and 1257 bp) were successfully fused with GUS reporter gene and integrated into rice plants, demonstrated that the promoter was functional in the heterologous plant system. The presence of blue GUS staining was observed on the leaf, root, stem, and grain of the transgenic rice regardless of the promoter length used and stayed functional up to the next generation. GUS qualitative analysis confirmed that the shorter promoter length generated a stronger GUS activity in comparison to the longer one. This indicated that the presence of repressor elements in between the -2771 bp and -1257 bp regions. The preliminary results shed light on the strong promoter activity in the rice endosperm tissue. It can become an alternative to the collection of plant promoters that can be used for grain quality improvement and biofortification.

scholarly journals PHLOEM PROMOTERS IN TRANSGENIC SWEET ORANGE ARE DIFFERENTIALLY TRIGGERED BY Candidatus Liberibacter asiaticus

2017 ◽  
Vol 39 (3) ◽  
Author(s):  
LUZIA YURIKO MIYATA ◽  
RICARDO HARAKAVA ◽  
LÍSIA BORGES ATTÍLIO ◽  
BEATRIZ MADALENA JANUZZI MENDES ◽  
JOÃO ROBERTO SPOTTI LOPES ◽  
...  
Keyword(s):  
Reporter Gene ◽  
Citrus Sinensis ◽  
Transgene Expression ◽  
Sweet Orange ◽  
Candidatus Liberibacter Asiaticus ◽  
Specific Expression ◽  
Gus Reporter ◽  
Uida Expression ◽  
Candidatus Liberibacter ◽  
Liberibacter Asiaticus

ABSTRACT The use of promoters preferentially expressed in specific plant tissues is a desirable strategy to search for resistance for pathogens that colonize these tissues. The bacterium Candidatus Liberibacter asiaticus (Las), associated with huanglongbing disease (HLB) of citrus, colonizes phloem vessels. Some promoters, besides conferring tissue-specific expression, can also respond to the presence of the pathogen. The objective of the present study was to verify if the presence of Las could modulate the activation of the phloem-specific promoters AtPP2 (Arabidopsis thaliana phloem protein 2), AtSUC2 (A. thaliana sucrose transporter 2) and CsPP2 ( pCitrus phloemrotein 2), known to be expressed in Citrus sinensis phloem. ‘Hamlin’ sweet orange plants (Citrus sinensis L. Osbeck) transformed with the uidA (GUS) reporter gene under the control of AtPP2, AtSUC2 and CsPP2 promoters were infected to evaluate the interdependence between transgene expression and the concentration of Las. Plants were inoculated with Las by Diaphorina citri and eighteen months later, bacterial concentration and uidA expression were determined by qPCR and RT-qPCR, respectively. Reporter gene expression driven by AtSUC2 promoter was strongly and positively correlated with Las concentration. Therefore, this promoter combines desirable features of both tissue-specificity and pathogen-inducibility for the production of transgenic plants tolerant to Las.

Matrix attachment regions and regulated transcription increase and stabilize transgene expression

2005 ◽  
Vol 3 (5) ◽  
pp. 535-543 ◽  
Author(s):  
Rita Abranches ◽  
Randall W. Shultz ◽  
William F. Thompson ◽  
George C. Allen
Keyword(s):  
Transgene Expression ◽  
Matrix Attachment Regions

scholarly journals Overexpression of the pitaya phosphoethanolamine N-methyltransferase gene (HpPEAMT) enhanced simulated drought stress in tobacco

2021 ◽  
Author(s):  
Ai-Hua Wang ◽  
Lan Yang ◽  
Xin-Zhuan Yao ◽  
Xiao-Peng Wen
Keyword(s):  
Drought Stress ◽  
Stress Response ◽  
Transgenic Plants ◽  
Drought Tolerance ◽  
Transgenic Tobacco ◽  
Sequence Similarity ◽  
Amino Acid Sequence Similarity ◽  
Transgenic Tobacco Plants ◽  
Wild Type ◽  
Tobacco Plants

AbstractPhosphoethanolamine N-methyltransferase (PEAMTase) catalyzes the methylation of phosphoethanolamine to produce phosphocholine and plays an important role in the abiotic stress response. Although the PEAMT genes has been isolated from many species other than pitaya, its role in the drought stress response has not yet been fully elucidated. In the present study, we isolated a 1485 bp cDNA fragment of HpPEAMT from pitaya (Hylocereus polyrhizus). Phylogenetic analysis showed that, during its evolution, HpPEAMT has shown a high degree of amino acid sequence similarity with the orthologous genes in Chenopodiaceae species. To further investigate the function of HpPEAMT, we generated transgenic tobacco plants overexpressing HpPEAMT, and the transgenic plants accumulated significantly more glycine betaine (GB) than did the wild type (WT). Drought tolerance trials indicated that, compared with those of the wild-type (WT) plants, the roots of the transgenic plants showed higher drought tolerance ability and exhibited improved drought tolerance. Further analysis revealed that overexpression of HpPEAM in Nicotiana tabacum resulted in upregulation of transcript levels of GB biosynthesis-related genes (NiBADH, NiCMO and NiSDC) in the leaves. Furthermore, compared with the wild-type plants, the transgenic tobacco plants displayed a significantly lower malondialdehyde (MDA) accumulation and higher activities of the superoxide dismutase (SOD) and peroxidase (POD) antioxidant enzymes under drought stress. Taken together, our results suggested that HpPEAMT enhanced the drought tolerance of transgenic tobacco.

scholarly journals The Promoter of the Rat Gonadotropin-Releasing Hormone Receptor Gene Directs the Expression of the Human Placental Alkaline Phosphatase Reporter Gene in Gonadotrope Cells in the Anterior Pituitary Gland as well as in Multiple Extrapituitary Tissues

Endocrinology ◽  
2004 ◽  
Vol 145 (2) ◽  
pp. 983-993 ◽  
Author(s):  
Anne Granger ◽  
Valérie Ngô-Muller ◽  
Christian Bleux ◽  
Céline Guigon ◽  
Hanna Pincas ◽  
...  
Keyword(s):  
Alkaline Phosphatase ◽  
Reporter Gene ◽  
Transgene Expression ◽  
Pituitary Cells ◽  
Placental Alkaline Phosphatase ◽  
R Gene ◽  
Glycoprotein Hormone ◽  
Steroidogenic Factor 1 ◽  
Functional Sites ◽  
Human Placental Alkaline Phosphatase

Abstract Previous studies dealing with the mechanisms underlying the tissue-specific and regulated expression of the GnRH receptor (GnRH-R) gene led us to define several cis-acting regulatory sequences in the rat GnRH-R gene promoter. These include functional sites for steroidogenic factor 1, activator protein 1, and motifs related to GATA and LIM homeodomain response elements as demonstrated primarily in transient transfection assays in mouse gonadotrope-derived cell lines. To understand these mechanisms in more depth, we generated transgenic mice bearing the 3.3-kb rat GnRH-R promoter linked to the human placental alkaline phosphatase reporter gene. Here we show that the rat GnRH-R promoter drives the expression of the reporter gene in pituitary cells expressing the LHβ and/or FSHβ subunit but not in TSHβ- or GH-positive cells. Furthermore, the spatial and temporal pattern of the transgene expression during the development of the pituitary was compatible with that characterizing the emergence of the gonadotrope lineage. In particular, transgene expression is colocalized with the expression of the glycoprotein hormone α-subunit at embryonic day 13.5 and with that of steroidogenic factor 1 at later stages of pituitary development. Transgene expression was also found in specific brain areas, such as the lateral septum and the hippocampus. A single promoter is thus capable of directing transcription in highly diverse tissues, raising the question of the different combinations of transcription factors that lead to such a multiple, but nevertheless cell-specific, expressions of the GnRH-R gene.

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