Cloning and Analysis of Vacuolar Invertase Gene (MeVINV2) Promoter from Cassava (Manihot Esculenta Crantz)

2014 ◽  
Vol 618 ◽  
pp. 259-263
Author(s):  
Jiao Liu ◽  
Yan Ping Hu ◽  
Wen Rui Xia ◽  
Yuan Yao ◽  
Yang Zhou ◽  
...  
Keyword(s):  
Gene Promoter ◽  
Pcr Amplification ◽  
Promoter Sequence ◽  
Regulatory Elements ◽  
Starch Accumulation ◽  
Manihot Esculenta Crantz ◽  
Vacuolar Invertase ◽  
Biological Stress ◽  
Invertase Gene ◽  
Vacuolar Invertases

Vacuolar invertases play a vital role in the progress of cassava tuber roots starch accumulation. In order to study the regulating mechanism of cassava vacuolar invertases, the promoter of cassava vacuolar invertase 2 (MeVINV2) was isolated using the PCR amplification approach, starting with a part of coding sequences. Sequencing result showed that 47 bp MeVINV2 gene CDS sequence and 1242 bp potential promoter sequence was obtained. PlantCARE analysis revealed that the MeVINV2 gene promoter contains typical eukaryotic elements CAAT box and TATA box, and also several light-responsive elements and stress-responsive elements. These cis-acting regulatory elements might be associated to the vacuolar invertase gene function of cassava starch accumulation and biological stress defense.

Cloning and Analysis of MeCWINV6 Promoter from Biofuel Plant Cassava (Manihot esculenta Crantz)

2014 ◽  
Vol 986-987 ◽  
pp. 25-29
Author(s):  
Jiao Liu ◽  
Wen Rui Xia ◽  
Yan Ping Hu ◽  
Yuan Yao ◽  
Shao Ping Fu ◽  
...  
Keyword(s):  
Cell Wall ◽  
Pcr Amplification ◽  
Promoter Sequence ◽  
Starch Accumulation ◽  
Cell Wall Invertase ◽  
Potential Promoter ◽  
Invertase Gene ◽  
Responsive Element ◽  
Source Sink ◽  
Insight Into

In order to gain insight into the specific function of the cassava cell wall invertase 6 (MeCWINV6), the promoter sequence of MeCWINV6 gene was cloned using the PCR amplification approach. 118 bp CDS sequence and 1042 bp potential promoter sequence of MeCWINV6 gene were obtained. PlantCARE analyzed the putative cis-elements in silico revealed that these elements can be grouped into five classes: basic transcription elements (CAAT box and TATA box), light responsive elements (ACE, AE-box, ATCT-motif, AT1-motif, Box 4, GAG-motif, GT1-motif and Sp1), phytohormone responsive motifs (GARE-motif, TATC-box, TGACG-motif and TCA-element), defense and stress responsive element (TC-rich repeats and HSE), wounding and pathogen responsive elements (W-box and WUN-motif). This data demonstrate that it might be associated to regulate the cell wall invertase gene function in source-sink relations of cassava starch accumulation and response to internal and environmental stimuli.

scholarly journals Arabidopsis thaliana Myb59 Gene Is Involved in the Response to Heterodera schachtii Infestation, and Its Overexpression Disturbs Regular Development of Nematode-Induced Syncytia

2021 ◽  
Vol 22 (12) ◽  
pp. 6450
Author(s):  
Anita Wiśniewska ◽  
Kamila Wojszko ◽  
Elżbieta Różańska ◽  
Klaudia Lenarczyk ◽  
Karol Kuczerski ◽  
...  
Keyword(s):  
Cell Metabolism ◽  
Gene Promoter ◽  
Promoter Sequence ◽  
Regulatory Elements ◽  
Heterodera Schachtii ◽  
Myb Transcription Factor ◽  
Parasitic Nematodes ◽  
Regulatory Sequences ◽  
Gene Encoding ◽  
Constitutive Overexpression

Transcription factors are proteins that directly bind to regulatory sequences of genes to modulate and adjust plants’ responses to different stimuli including biotic and abiotic stresses. Sedentary plant parasitic nematodes, such as beet cyst nematode, Heterodera schachtii, have developed molecular tools to reprogram plant cell metabolism via the sophisticated manipulation of genes expression, to allow root invasion and the induction of a sequence of structural and physiological changes in plant tissues, leading to the formation of permanent feeding sites composed of modified plant cells (commonly called a syncytium). Here, we report on the AtMYB59 gene encoding putative MYB transcription factor that is downregulated in syncytia, as confirmed by RT-PCR and a promoter pMyb59::GUS activity assays. The constitutive overexpression of AtMYB59 led to the reduction in A. thaliana susceptibility, as indicated by decreased numbers of developed females, and to the disturbed development of nematode-induced syncytia. In contrast, mutant lines with a silenced expression of AtMYB59 were more susceptible to this parasite. The involvement of ABA in the modulation of AtMYB59 gene transcription appears feasible by several ABA-responsive cis regulatory elements, which were identified in silico in the gene promoter sequence, and experimental assays showed the induction of AtMYB59 transcription after ABA treatment. Based on these results, we suggest that AtMYB59 plays an important role in the successful parasitism of H. schachtii on A. thaliana roots.

scholarly journals Cloning and Functional Analysis of Rat Tweety-Homolog 1 Gene Promoter

2021 ◽  
Author(s):  
Malgorzata Gorniak-Walas ◽  
Karolina Nizinska ◽  
Katarzyna Lukasiuk
Keyword(s):  
Transcriptional Regulation ◽  
Reporter Gene ◽  
Hippocampal Neurons ◽  
Gene Promoter ◽  
Promoter Sequence ◽  
Regulatory Elements ◽  
Luciferase Reporter ◽  
Luciferase Reporter Gene ◽  
Gene Assay

AbstractTweety-homolog 1 protein (Ttyh1) is abundantly expressed in neurons in the healthy brain, and its expression is induced under pathological conditions. In hippocampal neurons in vitro, Ttyh1 was implicated in the regulation of primary neuron morphology. However, the mechanisms that underlie transcriptional regulation of the Ttyh1 gene in neurons remain elusive. The present study sought to identify the promoter of the Ttyh1 gene and functionally characterize cis-regulatory elements that are potentially involved in the transcriptional regulation of Ttyh1 expression in rat dissociated hippocampal neurons in vitro. We cloned a 592 bp rat Ttyh1 promoter sequence and designed deletion constructs of the transcription factors specificity protein 1 (Sp1), E2F transcription factor 3 (E2f3), and achaete-scute homolog 1 (Ascl1) that were fused upstream of a luciferase reporter gene in pGL4.10[luc2]. The luciferase reporter gene assay showed the possible involvement of Ascl1, Sp1, and responsive cis-regulatory elements in Ttyh1 expression. These findings provide novel information about Ttyh1 gene regulation in neurons.

scholarly journals CLONING OF ProAV PROMOTER ISOLATED FROM TIGER PRAWN Penaeus monodon

2009 ◽  
Vol 4 (1) ◽  
pp. 1
Author(s):  
Andi Parenrengi ◽  
Alimuddin Alimuddin ◽  
Sukenda Sukenda ◽  
Komar Sumantadinata ◽  
Muhammad Yamin ◽  
...  
Keyword(s):  
Regulation Of Gene Expression ◽  
Penaeus Monodon ◽  
Gene Promoter ◽  
Promoter Sequence ◽  
Regulatory Elements ◽  
Polymerase Chain Reaction Method ◽  
Viral Gene ◽  
Isolation And Characterization ◽  
Tiger Prawn

Promoter is a specific DNA sequence involved in the transcription of a particular gene. It is usually located in the upstream of the gene they regulate. Isolation and characterization of promoter is essentially needed in order to establish the sequence analysis and transcription factor that are used in the regulation of gene expression. The research was conducted to analyze the characteristics of Penaeus monodon anti viral gene promoter (ProAV) towards generation of auto-transgenic tiger prawn, P. monodon. ProAV promoter was isolated by PCR (Polymerase Chain Reaction) method and the purified DNA fragment was cloned into pGEM-T Easy cloning vector. The promoter sequence was characterized by using BLAST-N and Genetyx version 7 softwares. The results showed the success in isolating a promoter from tiger prawn of 368 bp in length. BLAST-N analysis showed that the sequence of isolated promoter has high similarity (95%-98%) compared to the other promoters in the GeneBank. The study revealed the existence of important transcription factors (TATA box, MRE, TCF-1, and other potential regulatory elements) are identified in the promoter sequence.

ALLELE DIVERSITY FOR ACID VACUOLAR INVERTASE GENE Pain-1 FRAGMENT IN POTATO (Solanum tuberosum L.) VARIETIES AND LINES

2018 ◽  
Vol 53 (1) ◽  
pp. 132-139
Author(s):  
M.A. Slugina ◽  
◽  
E.O. Shmelkova ◽  
A.A. Meleshin ◽  
E.Z. Kochieva ◽  
...  
Keyword(s):  
Solanum Tuberosum ◽  
Solanum Tuberosum L ◽  
Vacuolar Invertase ◽  
Invertase Gene

scholarly journals Suppression of the Vacuolar Invertase Gene Prevents Cold-Induced Sweetening in Potato

2010 ◽  
Vol 154 (2) ◽  
pp. 939-948 ◽  
Author(s):  
Pudota B. Bhaskar ◽  
Lei Wu ◽  
James S. Busse ◽  
Brett R. Whitty ◽  
Andy J. Hamernik ◽  
...  
Keyword(s):  
Vacuolar Invertase ◽  
Invertase Gene ◽  
Cold Induced Sweetening

Control of mRNA turnover as a mechanism of glucose repression in Saccharomyces cerevisiae

1992 ◽  
Vol 12 (7) ◽  
pp. 2941-2948
Author(s):  
A Lombardo ◽  
G P Cereghino ◽  
I E Scheffler
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Succinate Dehydrogenase ◽  
Half Life ◽  
Glucose Repression ◽  
Promoter Sequence ◽  
Regulatory Elements ◽  
Mrna Levels ◽  
Protein Subunit ◽  
Gene Encoding ◽  
Regulatory Complex

We have examined the expression of the gene encoding the iron-protein subunit (Ip) of succinate dehydrogenase in Saccharomyces cerevisiae. The gene had been cloned by us and shown to be subject to glucose regulation (A. Lombardo, K. Carine, and I. E. Scheffler, J. Biol. Chem. 265:10419-10423, 1990). We discovered that a significant part of the regulation of the Ip mRNA levels by glucose involves the regulation of the turnover rate of this mRNA. In the presence of glucose, the half-life appears to be less than 5 min, while in glycerol medium, the half-life is greater than 60 min. The gene is also regulated transcriptionally by glucose. The upstream promoter sequence appeared to have four regulatory elements with consensus sequences shown to be responsible for the interaction with the HAP2/3/4 regulatory complex. A deletion analysis has shown that the two distal elements are redundant. These measurements were carried out by Northern (RNA) analyses of Ip mRNA transcripts as well as by assays of beta-galactosidase activity in cells carrying constructs of the Ip promoter linked to the lacZ coding sequence. These observations on the regulation of mRNA stability were also extended to the mRNA of the flavoprotein subunit of succinate dehydrogenase and in some experiments of iso-1-cytochrome c.

The Barley HvSTP13GR mutant triggers resistance against biotrophic fungi

2021 ◽  
Author(s):  
Caroline Ines Skoppek ◽  
Wilko Punt ◽  
Marleen Heinrichs ◽  
Frank Ordon ◽  
Gwendolin Wehner ◽  
...  
Keyword(s):  
Plant Protection ◽  
Rust Fungus ◽  
Durable Resistance ◽  
Sugar Transport ◽  
Gene Promoter ◽  
Monosaccharide Transporter ◽  
Base Editing ◽  
Biological Stress ◽  
Biotrophic Fungi ◽  
Promoter Studies

High-yielding and stress resistant crops are essential to ensure future food supply. Barley is an important crop to feed livestock and to produce malt, but the annual yield is threatened by pathogen infections. Pathogens can trigger an altered sugar partitioning in the host plant, that possibly leads to an advantage for the pathogen. Hampering these processes represents a promising strategy to potentially increase resistance. We analyzed the response of the barley monosaccharide transporter HvSTP13 towards biotic stress and its potential use for plant protection. The expression of HvSTP13 increased upon bacterial and fungal PAMP application, suggesting a PAMP-triggered signaling that converged on the transcriptional induction of the gene. Promoter studies indicate a region that is likely targeted by transcription factors downstream of PAMP-triggered immunity pathways. We confirmed that the non-functional HvSTP13GR variant confers resistance against an economically relevant biotrophic rust fungus, in barley. In addition, we established targeted CRISPR/Cas9 cytosine base editing in barley protoplasts to generate alternative HvSTP13 mutants and characterized the sugar transport activity and subcellular localization of the proteins. These mutants represent promising variants for future resistance analysis. Our experimental setup provides basal prerequisites to further decode the role of HvSTP13 in response to biological stress. Moreover, in line with other studies, our experiments indicate that the alteration of sugar partitioning pathways, in a host pathogen interaction, is a promising approach to achieve broad and durable resistance in plants.

scholarly journals Brain and muscle creatine kinase genes contain common TA-rich recognition protein-binding regulatory elements.

1990 ◽  
Vol 10 (9) ◽  
pp. 4826-4836 ◽  
Author(s):  
R A Horlick ◽  
G M Hobson ◽  
J H Patterson ◽  
M T Mitchell ◽  
P A Benfield
Keyword(s):  
Creatine Kinase ◽  
Binding Site ◽  
Regulatory Element ◽  
Gene Promoter ◽  
Regulatory Elements ◽  
Muscle Creatine Kinase ◽  
Enhancer Activity ◽  
L6 Myotubes ◽  
Recognition Protein ◽  
Muscle Creatine

We have previously reported that the rat brain creatine kinase (ckb) gene promoter contains an AT-rich sequence that is a binding site for a protein called TARP (TA-rich recognition protein). This AT-rich segment is a positively acting regulatory element for the ckb promoter. A similar AT-rich DNA segment is found at the 3' end of the 5' muscle-specific enhancer of the rat muscle creatine kinase (ckm) gene and has been shown to be necessary for full muscle-specific enhancer activity. In this report, we show that TARP binds not only to the ckb promoter but also to the AT-rich segment at the 3' end of the muscle-specific ckm enhancer. A second, weaker TARP-binding site was identified in the ckm enhancer and lies at the 5' end of the minimal enhancer segment. TARP was found in both muscle cells (C2 and L6 myotubes) and nonmuscle (HeLa) cells and appeared to be indistinguishable from both sources, as judged by gel retardation and footprinting assays. The TARP-binding sites in the ckm enhancer and the ckb promoter were found to be functionally interchangeable. We propose that TARP is active in both muscle and nonmuscle cells and that it is one of many potential activators that may interact with muscle-specific regulators to determine the myogenic phenotype.

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