A suite of novel promoters and terminators for plant biotechnology

2003 ◽  
Vol 30 (4) ◽  
pp. 443 ◽  
Author(s):  
Petra H. D. Schünmann ◽  
Danny J. Llewellyn ◽  
Brian Surin ◽  
Petra Boevink ◽  
Robert C. De Feyter ◽  
...  
Keyword(s):  
Solanum Tuberosum L ◽  
Marker Gene ◽  
Plant Biotechnology ◽  
Subterranean Clover ◽  
Expression Vectors ◽  
Crop Species ◽  
Specific Expression ◽  
Circular Dna ◽  
Subterranean Clover Stunt Virus ◽  
Gene Assay

The gene regulation signals from subterranean clover stunt virus (SCSV) were investigated for their expression in dicot plants. The SCSV genome has at least eight circular DNA molecules. Each circular DNA component contains a promoter element, a single open reading frame and a terminator. The promoters from seven of the segments were examined for their strength and tissue specificity in transgenic tobacco (Nicotiana tabacum L.), potato (Solanum tuberosum L.) and cotton (Gossypium hirsutum L.) using a GUS reporter gene assay system. While the promoters of many of the segments were poorly expressed, promoters derived from segments 4 and 7 were shown to direct high levels of expression in various plant tissues and organs. The segment 1 promoter directs predominantly callus-specific expression and, when used to control a selectable marker gene, facilitated the transformation of all three species (tobacco, potato and cotton). From the results, a suite of plant expression vectors (pPLEX) derived from the SCSV genome were constructed and used here to produce herbicide- and insect-resistant cotton, demonstrating their utility in the expression of foreign genes in dicot crop species and their potential for use in agricultural biotechnology.

A suite of novel promoters and terminators for plant biotechnology. II. The pPLEX series for use in monocots

2003 ◽  
Vol 30 (4) ◽  
pp. 453 ◽  
Author(s):  
Petra H.D. Schünmann ◽  
Brian Surin ◽  
Peter M. Waterhouse
Keyword(s):  
Transgene Expression ◽  
Expression Patterns ◽  
Plant Biotechnology ◽  
Leaf Tissue ◽  
Subterranean Clover ◽  
Expression Vectors ◽  
Reading Frame ◽  
Subterranean Clover Stunt Virus ◽  
Maize Ubiquitin Promoter ◽  
Low Levels

A suite of plant expression vectors (pPLEX), constructed from the gene regulation signals from subterranean clover stunt virus (SCSV) genome, has previously been used in dicot transformation for a variety of applications in plant biotechnology. To assess their use for the transformation of monocots, a number of modifications were made to the basic vector series and assessed in rice. In their unmodified forms, the SCSV promoters directed low levels of gene expression, however, insertion of an intron between the promoter and the transgene open reading frame (analogous to the rice actin and maize ubiquitin promoter systems) increased transgene expression 50-fold. The expression patterns from the intron-modified SCSV (segments 4 and 7) promoters were very similar to those directed by the actin or ubiquitin promoters. All promoter systems investigated directed expression that appeared to be constitutive within leaf tissue, and localised to the epidermal and vascular tissues of the root. The pPLEX vectors described here are an important counterpart to the dicot pPLEX series and have the potential to be useful in monocot research and biotechnology.

Cis/transgene optimization: systematic discovery of some key gene expression elements integrating bioinformatics and computational biology

2016 ◽  
Author(s):  
Saeid Kadkhodaei ◽  
Farahnaz S. Golestan Hashemi ◽  
Morvarid Akhavan Rezaei ◽  
Sahar Abbasiliasi ◽  
Tan Joo Shun ◽  
...  
Keyword(s):  
Gene Expression ◽  
Large Scale ◽  
Computational Analysis ◽  
Production Systems ◽  
Marker Gene ◽  
Optimization Procedure ◽  
Expression Vectors ◽  
Secretion Systems ◽  
Specific Expression ◽  
Gene Optimization

In recombinant protein production, quantity and quality are the major challenges particularly for large scale and high-throughput production systems. The present study mainly focused on computational analysis and in silico systematic discovery of some key functional gene expression elements in microalgae Dunaliella salina as a case study which there is no or poor information in this regard. Among the key factors, we took a shot at matrix attachment regions (MARs), translation initiation sites (TIS), signal peptide (SP) sequences, gene optimization and transformation system. Computational analysis of MARs sequences provided enough information about the structure of these sequences and led us to design an artificial MAR sequence considering the essential motifs and underlain rules. As the consensus TIS, we revealed that A-3, G-6C-5C-4 and G+1C+2G+3 arrange the specific context in this microalgae which help in locating the ribosome at the correct reading frame. Bioinformatics studies unveiled the sequence of MASTRAPLLALLALLCAGSARA with the highest signal score as the specific SP for secretion systems. A multi-criteria optimization procedure was performed to redesign the coding sequence of the BAR selectable marker gene. The optimized version of the gene mainly covered the host codon preference, the less structured mRNA and exposure of TIS. As the intragenic factors, we selected an efficient promoter, a 5ˈ-UTR and an intron from the closely related species (Chlamydomonas Sp.) to construct the specific expression vectors. The expression cassettes containing optimized genetic elements could be delivered into the microalgae cells and conferred the resistance to the transformants for at least 90 generations. The findings indicated that the MARs flanking the expression cassette along with the optimized expression elements particularly codon adaptation could potentially improve transformation efficiency and stability. The findings can be efficiently deployed as an empirical model for systematic discovery of the key expression elements and optimization of the cis/transgenes.

Invasion, development, growth and egg laying by Meloidogyne javanica in Brassicaceae crops

Nematology ◽  
2001 ◽  
Vol 3 (5) ◽  
pp. 463-472 ◽  
Author(s):  
John Kirkegaard ◽  
Rod McLeod ◽  
Christopher Steel
Keyword(s):  
Intermediate Host ◽  
Subterranean Clover ◽  
Meloidogyne Javanica ◽  
Mature Female ◽  
Egg Laying ◽  
Field Pea ◽  
Crop Species ◽  
Egg Masses ◽  
Brassicaceae Species ◽  
Host Status

AbstractInvasion, development and egg laying by Meloidogyne javanica in 11 Brassicaceae and four non-Brassicaceae crop species/subspecies was investigated. At 10 to15 and 15 to 20°C, fodder rape cv. Rangi was invaded less than the good hosts tomato cv. Grosse Lisse and field pea cv. Dun but more than the poor host oat cv. Cooba. With an inoculum of 50 second stage juveniles (J2), invasion of Rangi, and the intermediate host subterranean clover cv. Trikkala, were similarly invaded when inoculated with 50 and 100 J2, cv. Rangi was invaded less than tomato. The intermediate host subterranean clover cv. Trikkala and Rangi were similarly invaded when inoculated with 50 and 100 J2 but cv. Trikkala was less invaded with 200 J2. Oat cv. Cooba was always less invaded than the other hosts. Invasion of 3-week-old seedlings of cv. Rangi and 12 cultivars of seven other Brassicaceae crop species/subspecies were similar. Three weeks after inoculation, more M. javanica had developed to the mature female stage in tomato than in the eight Brassicaceae species/subspecies. Females growing in tomato and field pea were always larger than those in rape cv. Rangi. Females in Rangi were larger but those in oilseed radish cv. Adagio were smaller than in 11 other cultivars of seven Brassicaceae, except in plants grown in winter. Egg masses from four Brassicaceae species contained fewer eggs than egg masses from tomato at 6 weeks after inoculation, but at 7 and 8 weeks only those from fodder rape cv. Korina had consistently fewer than tomato. Results are discussed in relation to host status, glucosinolates and potential use of Brassicaceae for control of Meloidogyne.

Genetic patterns recognition in crop species using self-organizing map: the example of the highly heterozygous autotetraploid potato (Solanum tuberosum L.)

2020 ◽  
Vol 67 (4) ◽  
pp. 947-966
Author(s):  
M. C. Spanoghe ◽  
T. Marique ◽  
J. Rivière ◽  
M. Moulin ◽  
C. Dekuijper ◽  
...  
Keyword(s):  
Solanum Tuberosum ◽  
Solanum Tuberosum L ◽  
Self Organizing Map ◽  
Crop Species ◽  
Genetic Patterns ◽  
Self Organizing

scholarly journals Effect of single amino acid substitutions on the formation of the PlA and Bak alloantigenic epitopes

Blood ◽  
1991 ◽  
Vol 78 (3) ◽  
pp. 681-687 ◽  
Author(s):  
A Goldberger ◽  
M Kolodziej ◽  
M Poncz ◽  
JS Bennett ◽  
PJ Newman
Keyword(s):  
Amino Acid ◽  
Expression System ◽  
Single Amino Acid ◽  
Expression Vectors ◽  
Membrane Glycoproteins ◽  
Amino Acid Polymorphism ◽  
Specific Expression ◽  
Mammalian Expression ◽  
Single Amino Acid Polymorphisms ◽  
Necessary And Sufficient

Abstract The subunits that comprise the platelet-specific integrin alpha IIb beta 3 are polymorphic in nature, with several allelic forms present in the human gene pool. Minor changes in the secondary and tertiary structures of platelet membrane glycoproteins (GP) IIb and IIIa encoded by these alleles can result in an alloimmune reaction after transfusion or during pregnancy. To better understand the molecular structure of the PlA alloantigen system, located on GPIIIa, and the Bak alloantigen on GPIIb, we used a heterologous mammalian expression system to express these integrin subunits in their known polymorphic forms. An expression vector containing the PlA1 form of a GPIIIa cDNA, which encodes a leucine at amino acid 33 (Leu33), was modified to express the PlA2- associated form encoding a proline at amino acid 33 (Pro33). Similarly, a Baka GPIIb cDNA expressing an isoleucine at amino acid 843 (IIe843) was modified to express the Bakb form containing a serine at the same position (Ser843). Transfection of these vectors into COS cells resulted in the synthesis of GPIIb and GPIIIa molecules that were identical in size to those present in platelet lysates. Immunoprecipitation of the GPIIIa-transfected COS lysates with PlA)- specific alloantisera indicated that the Leu33 form was recognized only by anti-PIA1 sera while the Pro33 form was bound only by anti-PlA2 sera, showing that single amino acid polymorphisms are necessary and sufficient to direct the formation of the PlA1 and PlA2 alloepitopes. Similar experiments with Bak allele-specific expression vectors indicated that while the amino acid polymorphism (IIe843 in equilibrium Ser843) was necessary, posttranslational processing of pro-IIb was required for efficient exposure of both the Baka and Bakb alloepitopes.

Cloning of rat and mouse aquaporin-2 gene promoters and identification of a negative cis-regulatory element

1997 ◽  
Vol 273 (2) ◽  
pp. F264-F273 ◽  
Author(s):  
T. Rai ◽  
S. Uchida ◽  
F. Marumo ◽  
S. Sasaki
Keyword(s):  
Specific Binding ◽  
Regulatory Element ◽  
Gene Promoter ◽  
Nuclear Extract ◽  
Gene Promoters ◽  
Specific Expression ◽  
Cis Element ◽  
Aquaporin 2 ◽  
Gene Assay

The promoters of rat and mouse aquaporin-2 (AQP-2) genes were cloned and compared with that of human genes. Nucleotide identity up to -593 bp was 62%, and consensus sequences such as TATA box and adenosine 3',5'-cyclic monophosphate responsive element were conserved. Deoxyribonuclease I footprint assay revealed a footprinted region at -210 to -184 bp in rat AQP-2 gene promoter produced by nuclear extract from nonexpressing (liver) tissue. The sequence of this region included a GATA motif but otherwise showed no homology with any other previously known cis-elements. Electromobility shift assay and ultraviolet cross-linking analysis confirmed that specific binding proteins to this element were present in kidney, spleen, and liver and that these proteins were distinct from GATA factors. Both deletion and mutation of this cis-element abolished the protein DNA binding and increased promoter activity in in vitro reporter gene assay using rat cultured hepatocyte Ac2F cells, suggesting the negative regulatory role of this cis-element. These results indicate that tissue-specific expression of AQP-2 gene may in part be regulated by this novel negative acting cis-element.

PMI (manA) as a nonantibiotic selectable marker gene in plant biotechnology

2010 ◽  
Vol 105 (2) ◽  
pp. 141-148 ◽  
Author(s):  
P. Stoykova ◽  
P. Stoeva-Popova
Keyword(s):  
Selectable Marker ◽  
Marker Gene ◽  
Plant Biotechnology ◽  
Selectable Marker Gene

Varietal reaction and genetic resistance of subterranean clover (Trifolium subterraneum L.) to subterranean clover stunt virus infection

1960 ◽  
Vol 11 (5) ◽  
pp. 723 ◽  
Author(s):  
NW Grylls ◽  
JW Peak
Keyword(s):  
New Zealand ◽  
Virus Infection ◽  
Iberian Peninsula ◽  
Genetic Resistance ◽  
Subterranean Clover ◽  
Trifolium Subterraneum ◽  
Third Generation ◽  
Subterranean Clover Stunt Virus ◽  
Mediterranean Regions ◽  
The Mediterranean

Resistance to subterranean clover stunt virus was explored in 390 strains and named varieties of subterranean clover from the Mediterranean regions, England, France, the Iberian peninsula, New Zealand, and Australia. High levels of genetic resistance were shown in the Australian varieties Tallarook, Hill's Small, and Bass B. Resistance of a selected group of F2's was found to be midway between that of the parents. In selected groups of F4 generation hybrids, and in selected second and third generation backcrosses, resistance equal to that of Tallarook was shown. The apparent recovery of some plants during tests in the glass-house was shown to be a form of temporary tolerance to the virus.

scholarly journals Allele-specific expression identified rs2509956 as a novel long-distance cis-regulatory SNP for SCGB1A1, an important gene for multiple pulmonary diseases

2019 ◽  
Vol 317 (4) ◽  
pp. L456-L463
Author(s):  
Xiu-Xiong Li ◽  
Tao Peng ◽  
Jing Gao ◽  
Jia-Gang Feng ◽  
Dan-Dan Wu ◽  
...  
Keyword(s):  
Regulatory Elements ◽  
Pulmonary Diseases ◽  
Chronic Obstructive ◽  
Long Distance ◽  
Specific Expression ◽  
Allele Specific Expression ◽  
Chromosome Conformation ◽  
Functional Region ◽  
Gene Assay ◽  
Allele Specific

SCGB1A1 (secretoglobin family 1A member 1) is an important protein for multiple pulmonary diseases, especially asthma, chronic obstructive pulmonary disease, and lung cancer. One single-nucleotide polymorphism (SNP) at 5′-untranslated region of SCGB1A1, rs3741240, has been suggested to be associated with reduced protein expression and further asthma susceptibility. However, it was still unclear whether there were other cis-regulatory elements for SCGB1A1 that might further contribute to pulmonary diseases. Allele-specific expression (ASE) is a novel approach to identify the functional region in human genome. In the present study, we measured ASE on rs3741240 in lung tissues and observed a consistent excess of G allele over A ( P < 10−6), which indicated that this SNP or the one(s) in linkage disequilibrium (LD) could regulate SCGB1A1 expression. By analyzing 1000 Genomes Project data for Chinese, one SNP locating ~10.2 kb away and downstream of SCGB1A1, rs2509956, was identified to be in strong LD with rs3741240. Reporter gene assay confirmed that both SNPs could regulate gene expression in the lung cell. By chromosome conformation capture, it was verified that the region surrounding rs2509956 could interact with SCGB1A1 promoter region and act as an enhancer. Through chromatin immunoprecipitation and overexpression assay, the related transcription factor RELA (RELA proto-oncogene, NF-kB subunit) was recognized to bind the region spanning rs2509956. Our work identified a novel long-distance cis-regulatory SNP for SCGB1A1, which might contribute to multiple pulmonary diseases.

scholarly journals Transgenic Expression of Glucagon-Like Peptide-1 (GLP-1) and Activated Muscarinic Receptor (M3R) Significantly Improves Pig Islet Secretory Function

2017 ◽  
Vol 26 (5) ◽  
pp. 901-911 ◽  
Author(s):  
Nizar I. Mourad ◽  
Andrea Perota ◽  
Daela Xhema ◽  
Cesare Galli ◽  
Pierre Gianello
Keyword(s):  
Insulin Secretion ◽  
Muscarinic Receptor ◽  
Expression Vectors ◽  
Specific Expression ◽  
Transgenic Expression ◽  
Glucagon Like Peptide ◽  
Glucagon Like Peptide 1 ◽  
Glucose Stimulated Insulin Secretion ◽  
Type 3

Porcine islets show notoriously low insulin secretion levels in response to glucose stimulation. While this is somehow expected in the case of immature islets isolated from fetal and neonatal pigs, disappointingly low secretory responses are frequently reported in studies using in vitro-maturated fetal and neonatal islets and even fully differentiated adult islets. Herein we show that β-cell-specific expression of a modified glucagon-like peptide-1 (GLP-1) and of a constitutively activated type 3 muscarinic receptor (M3R) efficiently amplifies glucose-stimulated insulin secretion (GSIS). Both adult and neonatal isolated pig islets were treated with adenoviral expression vectors carrying sequences encoding for GLP-1 and/or M3R. GSIS from transduced and control islets was evaluated during static incubation and dynamic perifusion assays. While expression of GLP-1 did not affect basal or stimulated insulin secretion, activated M3R produced a twofold increase in both first and second phases of GSIS. Coexpression of GLP-1 and M3R caused an even greater increase in the secretory response, which was amplified fourfold compared to controls. In conclusion, our work highlights pig islet insulin secretion deficiencies and proposes concomitant activation of cAMP-dependent and cholinergic pathways as a solution to ameliorate GSIS from pig islets used for transplantation.

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