The first intron of rice EPSP synthase enhances expression of foreign gene

2003 ◽  
Vol 46 (6) ◽  
pp. 561 ◽  
Author(s):  
Junwang XU
Keyword(s):  
Foreign Gene ◽  
Epsp Synthase ◽  
First Intron

scholarly journals Targeted Deletion of the Wx Gene First Intron via CRISPR/Cas9 Significantly Increases Grain Amylose Content in Rice With a Wxb Allele, but Not in Rice With a Wxa Allele

2021 ◽  
Author(s):  
Xingdan Liu ◽  
Qi Ding ◽  
Wenshu Wang ◽  
Yanling Pan ◽  
Chao Tan ◽  
...  
Keyword(s):  
Gene Expression ◽  
Cooking Quality ◽  
Amylose Content ◽  
Mrna Level ◽  
Foreign Gene ◽  
Rice Grain ◽  
Wild Type ◽  
Targeted Deletion ◽  
First Intron ◽  
Eating And Cooking Quality

Abstract Background: Rice Waxy (Wx) gene plays a major role in seed amylose synthesis, and consequently controls grain amylose content. The expression of Wx gene is highly regulated at both transcriptional and post-transcriptional levels. Particularly, the GT/TT poplymorphism at the 5` splicing site of its 1st intron greatly affects this intron’s splicing efficiency and defines two predominant Wx alleles, Wxa and Wxb. Wxa rice often has intermediate to high amylose content, whereas Wxb rice has low to intermediate amylose content. A previous study indicates that rice Wx 1st intron significantly enhances gene expression when it is inserted into the 5` UTR (untranslated region) of a foreign gene. By deleting Wx 1st intron with the CRISPR/Cas9 technology, we intended to create a totally noval Wx allele, and further to investigate how the intron removal affects Wx gene expression and rice grain amylose content.Results: CRISPR/Cas9-mediated targeted deletion of Wx 1st intron was performed on 4 rice inbreds, KY131(Wxb), X32(Wxb), X35(Wxa) and X55(Wxlv). Complete deletion of the 1st intron occurred in 8.6%-11.8% of the primary transformants of these 4 inbreds. Transgene-free, homozygous mutants were obtained. Their grain amylose content and Wx gene expression were analyzed. Compared to the amylose content of wild type plants, mutants’ amylose content was significantly increased from 13.0% to about 24% in KY131 and X32 which both carried the Wxb allele. However, no significant differenece in aylose content was observed between wild type plants and mutants of X35 and X55 which carried the Wxa and Wxlv allele, respectively. Results of Wx gene expression analysis on wild type plants and mutants showed a high consistence with their amylose content results. Mutants of KY131 and X32 accumulated much more steady mRNA transcripts than their wild type plants, while steady mRNA level remained somehow unchanged between wild type plants and mutants of X35 and X55. Grain quality including appearance quality and ECQ(eating and cooking quality) that are tightly linked to amylose content was also evalued on wild type plants and mutants, and data were presented and analyzed. Conclusions:This study presents a novel and fast strategy to increase amylose content for rice inbreds carrying a Wxb allele. Our data strongly suggest that rice Wx 1st intron regulates Wx gene expression mainly at the post-transcriptinal level, not as previously thought that it influences Wx gene transcription as well. In addition, removal of the first intron creates a completely noval Wx allele. Further studies on this new Wx allele would provide invaluable insights into the regulation of Wx gene expression, which will help researchers to engineer more new alleles that leads to the breeding of rice cultivars with better eating and cooking quality.

scholarly journals Microinjection of Bovine Embryos with a Foreign Gene and Its Detection at the Blastocyst Stage

1993 ◽  
Vol 76 (11) ◽  
pp. 3392-3399 ◽  
Author(s):  
Esmail Behboodi ◽  
G.B. Anderson ◽  
S. Horvat ◽  
J.F. Medrano ◽  
J.D. Murray ◽  
...  
Keyword(s):  
Blastocyst Stage ◽  
Foreign Gene ◽  
Bovine Embryos

scholarly journals Engineering of Recombinant Sheep Pox Viruses Expressing Foreign Antigens

2021 ◽  
Vol 9 (5) ◽  
pp. 1005
Author(s):  
Olga Chervyakova ◽  
Elmira Tailakova ◽  
Nurlan Kozhabergenov ◽  
Sandugash Sadikaliyeva ◽  
Kulyaisan Sultankulova ◽  
...  
Keyword(s):  
Fluorescent Protein ◽  
Viral Vector ◽  
Foot And Mouth Disease ◽  
Open Reading Frames ◽  
Foreign Gene ◽  
Mouth Disease Virus ◽  
Foreign Genes ◽  
Green Fluorescent ◽  
Reading Frames

Capripoxviruses with a host range limited to ruminants have the great potential to be used as vaccine vectors. The aim of this work was to evaluate attenuated sheep pox virus (SPPV) vaccine strain NISKHI as a vector expressing several genes. Open reading frames SPPV020 (ribonucleotide kinase) and SPPV066 (thymidine kinase) were selected as sites for the insertion of foreign genes. Two integration plasmids with expression cassette were designed and constructed. Recombinant SPPVs expressing an enhanced green fluorescent protein (EGFP) (rSPPV(RRΔ)EGFP and rSPPV(TKΔ)EGFP), Foot-and-mouth disease virus capsid protein (VP1), and Brucella spp. outer membrane protein 25 (OMP25) (rSPPV(RRΔ)VP1A-(TKΔ)OMP25) were generated under the transient dominant selection method. The insertion of foreign genes into the SPPV020 and SPPV066 open reading frames did not influence the replication of the recombinant viruses in the cells. Successful foreign gene expression in vitro was assessed by luminescent microscopy (EGFP) and Western blot (VP1 and OMP25). Our results have shown that foreign genes were expressed by rSPPV both in permissive (lamb testicles) and non-permissive (bovine kidney, saiga kidney, porcine kidney) cells. Mice immunized with rSPPV(RRΔ)VP1A-(TKΔ)OMP25 elicited specific antibodies to both SPPV and foreign genes VP1 and OMP25. Thus, SPPV NISKHI may be used as a potential safe immunogenic viral vector for the development of polyvalent vaccines.

scholarly journals Genome-wide discovery of G-quadruplexes in barley

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
H. Busra Cagirici ◽  
Hikmet Budak ◽  
Taner Z. Sen
Keyword(s):  
Transcription Initiation ◽  
Large Window ◽  
First Intron ◽  
Domain Sequence ◽  
Genome Wide ◽  
Square Planar ◽  
Peak Points ◽  
Molecular Functions ◽  
Nucleic Acid Structures ◽  
Provided Examples

AbstractG-quadruplexes (G4s) are four-stranded nucleic acid structures with closely spaced guanine bases forming square planar G-quartets. Aberrant formation of G4 structures has been associated with genomic instability. However, most plant species are lacking comprehensive studies of G4 motifs. In this study, genome-wide identification of G4 motifs in barley was performed, followed by a comparison of genomic distribution and molecular functions to other monocot species, such as wheat, maize, and rice. Similar to the reports on human and some plants like wheat, G4 motifs peaked around the 5′ untranslated region (5′ UTR), the first coding domain sequence, and the first intron start sites on antisense strands. Our comparative analyses in human, Arabidopsis, maize, rice, and sorghum demonstrated that the peak points could be erroneously merged into a single peak when large window sizes are used. We also showed that the G4 distributions around genic regions are relatively similar in the species studied, except in the case of Arabidopsis. G4 containing genes in monocots showed conserved molecular functions for transcription initiation and hydrolase activity. Additionally, we provided examples of imperfect G4 motifs.

scholarly journals Future Prospects of Gene Therapy for Friedreich’s Ataxia

2021 ◽  
Vol 22 (4) ◽  
pp. 1815 ◽  
Author(s):  
Gabriel Ocana-Santero ◽  
Javier Díaz-Nido ◽  
Saúl Herranz-Martín
Keyword(s):  
Gene Therapy ◽  
Viral Vectors ◽  
Autosomal Recessive ◽  
State Of The Art ◽  
Friedreich’S Ataxia ◽  
Friedreich's Ataxia ◽  
First Intron ◽  
Progressive Neurodegeneration ◽  
Feasible Solutions ◽  
Neurogenetic Disease

Friedreich’s ataxia is an autosomal recessive neurogenetic disease that is mainly associated with atrophy of the spinal cord and progressive neurodegeneration in the cerebellum. The disease is caused by a GAA-expansion in the first intron of the frataxin gene leading to a decreased level of frataxin protein, which results in mitochondrial dysfunction. Currently, there is no effective treatment to delay neurodegeneration in Friedreich’s ataxia. A plausible therapeutic approach is gene therapy. Indeed, Friedreich’s ataxia mouse models have been treated with viral vectors en-coding for either FXN or neurotrophins, such as brain-derived neurotrophic factor showing promising results. Thus, gene therapy is increasingly consolidating as one of the most promising therapies. However, several hurdles have to be overcome, including immunotoxicity and pheno-toxicity. We review the state of the art of gene therapy in Friedreich’s ataxia, addressing the main challenges and the most feasible solutions for them.

scholarly journals Identification of an enhancer sequence within the first intron required for cartilage-specific transcription of the α2(XI) collagen gene. VOLUME 275 (2000) PAGES 12712-12718

2009 ◽  
Vol 284 (17) ◽  
pp. 11748.2-11748 ◽  
Author(s):  
Ying Liu ◽  
Haochuan Li ◽  
Kazuhiro Tanaka ◽  
Noriyuki Tsumaki ◽  
Yoshihiko Yamada
Keyword(s):  
Collagen Gene ◽  
Enhancer Sequence ◽  
First Intron

scholarly journals Mitochondrial Group II Introns, Cytochrome c Oxidase, and Senescence in Podospora anserina

1999 ◽  
Vol 19 (6) ◽  
pp. 4093-4100 ◽  
Author(s):  
Odile Begel ◽  
Jocelyne Boulay ◽  
Beatrice Albert ◽  
Eric Dufour ◽  
Annie Sainsard-Chanet
Keyword(s):  
Life Span ◽  
Integration Site ◽  
Group Ii Intron ◽  
Podospora Anserina ◽  
Wild Type ◽  
First Intron ◽  
Limited Life ◽  
Related Group ◽  
Group Ii ◽  
Mitochondrial Chromosome

ABSTRACT Podospora anserina is a filamentous fungus with a limited life span. It expresses a degenerative syndrome called senescence, which is always associated with the accumulation of circular molecules (senDNAs) containing specific regions of the mitochondrial chromosome. A mobile group II intron (α) has been thought to play a prominent role in this syndrome. Intron α is the first intron of the cytochrome c oxidase subunit I gene (COX1). Mitochondrial mutants that escape the senescence process are missing this intron, as well as the first exon of theCOX1 gene. We describe here the first mutant of P. anserina that has the α sequence precisely deleted and whose cytochrome c oxidase activity is identical to that of wild-type cells. The integration site of the intron is slightly modified, and this change prevents efficient homing of intron α. We show here that this mutant displays a senescence syndrome similar to that of the wild type and that its life span is increased about twofold. The introduction of a related group II intron into the mitochondrial genome of the mutant does not restore the wild-type life span. These data clearly demonstrate that intron α is not the specific senescence factor but rather an accelerator or amplifier of the senescence process. They emphasize the role that intron α plays in the instability of the mitochondrial chromosome and the link between this instability and longevity. Our results strongly support the idea that in Podospora, “immortality” can be acquired not by the absence of intron α but rather by the lack of active cytochromec oxidase.

Production of foreign proteins in the yeast Pichia pastoris

1995 ◽  
Vol 73 (S1) ◽  
pp. 891-897 ◽  
Author(s):  
James M. Cregg ◽  
David R. Higgins
Keyword(s):  
Gene Expression ◽  
Pichia Pastoris ◽  
Heterologous Gene Expression ◽  
Expression System ◽  
Methylotrophic Yeast ◽  
Culture Broth ◽  
Heterologous Gene ◽  
Foreign Gene ◽  
Heterologous Proteins ◽  
Yeast Pichia Pastoris

The methanol-utilizing yeast Pichia pastoris has been developed as a host system for the production of heterologous proteins of commercial interest. An industrial yeast selected for efficient growth on methanol for biomass generation, P. pastoris is readily grown on defined medium in continuous culture at high volume and density. A unique feature of the expression system is the promoter employed to drive heterologous gene expression, which is derived from the methanol-regulated alcohol oxidase I gene (AOX1) of P. pastoris, one of the most efficient and tightly regulated promoters known. The strength of the AOX1 promoter results in high expression levels in strains harboring only a single integrated copy of a foreign-gene expression cassette. Levels may often be further enhanced through the integration of multiple cassette copies into the P. pastoris genome and strategies to construct and select multicopy cassette strains have been devised. The system is particularly attractive for the secretion of foreign-gene products. Because P. pastoris endogenous protein secretion levels are low, foreign secreted proteins often appear to be virtually the only proteins in the culture broth, a major advantage in processing and purification. Key words: heterologous gene expression, methylotrophic yeast, Pichia pastoris, secretion, glycosylation.

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