scholarly journals FOREIGN GENE EXPRESSION IN TRANSGENIC CRANBERRY PLANTS

HortScience ◽  
1992 ◽  
Vol 27 (6) ◽  
pp. 621d-621
Author(s):  
Rod Serres ◽  
David Russell ◽  
Dan Mahr ◽  
Brent McCown
Keyword(s):  
Normal Growth ◽  
Foreign Gene ◽  
Foreign Gene Expression ◽  
Specific Expression ◽  
Genome Expression ◽  
Feeding Trials ◽  
Blackheaded Fireworm ◽  
Foreign Genes ◽  
Lepidopteran Resistance

Genetically transformed Vaccinium macrocarpon `Stevens' and `Pilgrim' plants have been obtained using electric discharge particle acceleration. Three foreign genes, kan encoding a selectable marker, gus a reporter gene, and B.t.k. conferring lepidopteran resistance, were incorporated into the genome. Expression of kan was assayed by culturing shoots in vitro on media with several concentrations of kanamycin. Expression among transformed clones (transclones) varied from high resistance (normal growth at 300 mg/L kan) to no resistance. Histochemical analyses for gus expression revealed variability among transclones. Some transclones exhibited no gus expression, others had consistent area-specific expression while others displayed random expression. In preliminary feeding trials with blackheaded fireworm larvae, B.t.k. expression was found to be ineffective at controlling insect development. We have recovered plants transformed with a different promoter driving the B.t.k. gene in an effort to enhance expression.

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 The ferroxidase LPR5 functions in the maintenance of phosphate homeostasis and is required for normal growth and development of rice

2020 ◽  
Vol 71 (16) ◽  
pp. 4828-4842
Author(s):  
Hao Ai ◽  
Yue Cao ◽  
Ajay Jain ◽  
Xiaowen Wang ◽  
Zhi Hu ◽  
...  
Keyword(s):  
Growth And Development ◽  
Xylem Sap ◽  
Sufficient Conditions ◽  
Normal Growth ◽  
Yield Potential ◽  
Specific Expression ◽  
Gus Reporter ◽  
Ferroxidase Activity ◽  
Vitro Analysis

Abstract Members of the Low Phosphate Root (LPR) family have been identified in rice (Oryza sativa) and expression analyses have been conducted. Here, we investigated the functions of one of the five members in rice, LPR5. qRT-PCR and promoter–GUS reporter analyses indicated that under Pi-sufficient conditions OsLPR5 was highly expressed in the roots, and specific expression occurred in the leaf collars and nodes, and its expression was increased under Pi-deficient conditions. In vitro analysis of the purified OsLPR5 protein showed that it exhibited ferroxidase activity. Overexpression of OsLPR5 triggered higher ferroxidase activity, and elevated concentrations of Fe(III) in the xylem sap and of total Fe in the roots and shoots. Transient expression of OsLPR5 in Nicotiana benthamiana provided evidence of its subcellular localization to the cell wall and endoplasmic reticulum. Knockout mutation in OsLPR5 by means of CRISPR-Cas9 resulted in adverse effects on Pi translocation, on the relative expression of Cis-NATOsPHO1;2, and on several morphological traits, including root development and yield potential. Our results indicate that ferroxidase-dependent OsLPR5 has both a broad-spectrum influence on growth and development in rice as well as affecting a subset of physiological and molecular traits that govern Pi homeostasis.

scholarly journals Coronaviruses as Vectors: Position Dependence of Foreign Gene Expression

2003 ◽  
Vol 77 (21) ◽  
pp. 11312-11323 ◽  
Author(s):  
Cornelis A. M. de Haan ◽  
Linda van Genne ◽  
Jeroen N. Stoop ◽  
Haukeline Volders ◽  
Peter J. M. Rottier
Keyword(s):  
Gene Expression ◽  
Expression Vectors ◽  
Luciferase Expression ◽  
Foreign Gene ◽  
Regulatory Sequences ◽  
Expression Levels ◽  
Genomic Position ◽  
Position Dependence ◽  
Foreign Genes

ABSTRACT Coronaviruses are the enveloped, positive-stranded RNA viruses with the largest RNA genomes known. Several features make these viruses attractive as vaccine and therapeutic vectors: (i) deletion of their nonessential genes is strongly attenuating; (ii) the genetic space thus created allows insertion of foreign information; and (iii) their tropism can be modified by manipulation of the viral spike. We studied here their ability to serve as expression vectors by inserting two different foreign genes and evaluating systematically the genomic position dependence of their expression, using a murine coronavirus as a model. Renilla and firefly luciferase expression cassettes, each provided with viral transcription regulatory sequences (TRSs), were inserted at several genomic positions, both independently in different viruses and combined within one viral genome. Recombinant viruses were generated by using a convenient method based on targeted recombination and host cell switching. In all cases high expression levels of the foreign genes were observed without severe effects on viral replication in vitro. The expression of the inserted gene appeared to be dependent on its genomic position, as well as on the identity of the gene. Expression levels increased when the luciferase gene was inserted closer to the 3′ end of the genome. The foreign gene insertions generally reduced the expression of upstream viral genes. The results are consistent with coronavirus transcription models in which the transcription from upstream TRSs is attenuated by downstream TRSs. Altogether, our observations clearly demonstrate the potential of coronaviruses as (multivalent) expression vectors.

Introduction of Foreign Genes into Silkworm Eggs by Electroporation and Its Application in Transgenic Vector Test

2004 ◽  
Vol 36 (5) ◽  
pp. 323-330 ◽  
Author(s):  
Xiu-Yang Guo ◽  
Liang Dong ◽  
Sheng-Peng Wang ◽  
Ting-Qing Guo ◽  
Jian-Yang Wang ◽  
...  
Keyword(s):  
Instar Larva ◽  
Foreign Gene ◽  
Positive Ratio ◽  
Significant Difference ◽  
Voltage Dependent ◽  
Localization Signal ◽  
Foreign Genes ◽  
The One ◽  
Transposase Mrna

Abstract Electroporation as a methodology to introduce foreign genes into silkworm eggs was systematically analyzed. The foreign gene in both the newly hatched and 3rd instar larva DNA can be detected by PCR. The amount of foreign gene in 3rd instar larva DNA was about 1/1000 of that in newly hatched larva DNA. The ratio of foreign gene entering into silkworm eggs was voltage dependent and showed significant difference between the tested silkworm strains. When the piggyBac transposon system was applied, the effect of nuclear localization signal (NLS) peptide and the in vitro transcribed transposase mRNA on the transposition rate has been measured. Results showed that the in vitro transcribed transposase mRNA facilitated transposition to take place earlier and NLS could result in higher transposition probability and earlier transposition as well. When linearized vectors containing varied length of flanking homologous sequences around a reporter gene were introduced into silkworm eggs by electroporation, the one with 2.6 kb total arm length gave higher G1 positive ratio than that with total arm length of 1.5 kb and 800 bp.

scholarly journals Foreign Gene Expression in the Gonads of Chimaeric Chicken Embryos by Transfer of Primordial Germ Cells Transfected in vitro by Lipofection for 24 Hours

2000 ◽  
Vol 71 (3) ◽  
pp. 308-311
Author(s):  
Mitsuru NAITO ◽  
Yuko MATSUBARA ◽  
Takashi HARUMI ◽  
Takahiro TAGAMI ◽  
Michiharu SAKURAI ◽  
...  
Keyword(s):  
Gene Expression ◽  
Germ Cells ◽  
Primordial Germ Cells ◽  
Foreign Gene ◽  
Foreign Gene Expression ◽  
Chicken Embryos

scholarly journals Rapid Delivery of Foreign Genes into Plants by Direct Rub-Inoculation with Intact Plasmid DNA of a Tomato Bushy Stunt Virus Gene Vector

1999 ◽  
Vol 73 (9) ◽  
pp. 7823-7829 ◽  
Author(s):  
Herman B. Scholthof
Keyword(s):  
Gene Expression ◽  
Tomato Bushy Stunt Virus ◽  
Cauliflower Mosaic Virus ◽  
35S Promoter ◽  
Foreign Gene ◽  
Gene Vector ◽  
Foreign Gene Expression ◽  
Foreign Genes ◽  
Nopaline Synthase Gene ◽  
Delta Virus

ABSTRACT Tomato bushy stunt virus (TBSV) cDNA, positioned between a modified cauliflower mosaic virus 35S promoter and the hepatitis delta virus antigenomic ribozyme with a downstream nopaline synthase gene polyadenylation signal, established infections upon rub-inoculation of plants with intact plasmids. Application of this methodology produced a TBSV DNA-based gene vector which yielded readily detectable levels of localized foreign gene expression in inoculated leaves. This is the first demonstration of an infectious DNA from a member of theTombusviridae which permits rapid TBSV-mediated foreign-gene expression upon direct rub-inoculation of miniprep DNA onto a variety of plant species.

scholarly journals In vivo manipulation of foreign gene expression by steroid administration in the oviduct of laying hens

1999 ◽  
Vol 163 (2) ◽  
pp. 173-179 ◽  
Author(s):  
HM Park ◽  
T Muramatsu
Keyword(s):  
Gene Expression ◽  
Laying Hens ◽  
Foreign Gene ◽  
Foreign Gene Expression ◽  
Response Elements ◽  
Steroid Administration ◽  
Steroid Response ◽  
In Vivo Induction

The experiments described herein were conducted to examine whether or not steroid administration allows in vivo induction of foreign gene expression in the oviduct of laying hens. The chloramphenicol acetyltransferase reporter gene driven by several viral and cellular promoters with or without steroid response elements was transfected by in vivo electroporation. The results indicated that in vivo, as observed in vitro, steroid administration induced transcriptional activities of the promoters with steroid response elements but it did not do so without steroid response elements. Our data implicate, therefore, that in vivo induction of foreign gene expression is possible in the oviduct of laying hens, and that the present in vivo gene transfer approach would serve as a useful tool to elucidate the mechanism of tissue-specific and steroid-induced transcription of chicken egg white genes.

scholarly journals FOREIGN GENE TRANSFER TO SWEET POTATO (IPOMOEA BATATAS)

HortScience ◽  
1991 ◽  
Vol 26 (5) ◽  
pp. 492f-492 ◽  
Author(s):  
C.S. Prakash ◽  
U. Varadarajan ◽  
A. S. Kumar
Keyword(s):  
Gene Transfer ◽  
Sweet Potato ◽  
Ipomoea Batatas ◽  
Microprojectile Bombardment ◽  
Pcr Amplification ◽  
Marker Genes ◽  
Foreign Gene ◽  
Quantitative Expression ◽  
Foreign Genes

Development of a gene transfer system will enable rapid introduction of agronomically useful genes into elite cultivars of sweet potato. We compared microprojectile bombardment and Agrobacterium cocultivation approaches to introduce foreign genes into the genome of two sweet potato cultivars. Chimeric marker genes (gusA and kan) were successfully introduced into cvs. Jewel and TIS-70357 using both approaches. However, transgenic plants were generated in vitro using only the Agrobacterium approach. Callus and root isolates with stable expression of gusA gene were obtained using the microprojectile method. Expression of the screenable marker gusA gene was detected by histochemical assays. Integration of the introduced gene into the genome of sweet potato was confirmed by polymerase chain reaction (PCR) amplification of the kan gene and Southern blot analyses. Transgenic sweet potato plants from two cultivars are being raised and studied for quantitative expression and localization of the introduced genes. These results show that foreign genes can be successfully introduced and expressed in sweet potato. Current efforts are directed at optimizing several variables to increase the transformation efficiencies and to generate transgenic cultivars with foreign genes of agricultural importance.

Probing the 14-3-3 Isoform-Specificity Profile of Interactions Stabilized by Fusicoccin A

2020 ◽  
Author(s):  
James Frederich ◽  
Ananya Sengupta ◽  
Josue Liriano ◽  
Ewa A. Bienkiewicz ◽  
Brian G. Miller
Keyword(s):  
Protein Interactions ◽  
Neurological Diseases ◽  
Adapter Proteins ◽  
Protein Protein Interactions ◽  
Specific Expression ◽  
Individual Client ◽  
Isoform Specificity ◽  
Fusicoccin A

Fusicoccin A (FC) is a fungal phytotoxin that stabilizes protein–protein interactions (PPIs) between 14-3-3 adapter proteins and their phosphoprotein interaction partners. In recent years, FC has emerged as an important chemical probe of human 14-3-3 PPIs implicated in cancer and neurological diseases. These previous studies have established the structural requirements for FC-induced stabilization of 14-3-3·client phosphoprotein complexes; however, the effect of different 14-3-3 isoforms on FC activity has not been systematically explored. This is a relevant question for the continued development of FC variants because there are seven distinct isoforms of 14-3-3 in humans. Despite their remarkable sequence and structural similarities, a growing body of experimental evidence supports both tissue-specific expression of 14-3-3 isoforms and isoform-specific functions <i>in vivo</i>. Herein, we report the isoform-specificity profile of FC <i>in vitro</i>using recombinant human 14-3-3 isoforms and a focused library of fluorescein-labeled hexaphosphopeptides mimicking the C-terminal 14-3-3 recognition domains of client phosphoproteins targeted by FC in cell culture. Our results reveal modest isoform preferences for individual client phospholigands and demonstrate that FC differentially stabilizes PPIs involving 14-3-3s. Together, these data provide strong motivation for the development of non-natural FC variants with enhanced selectivity for individual 14-3-3 isoforms.

Non-Viral Vectors for Gene Delivery

2018 ◽  
Vol 9 (1) ◽  
pp. 4-11 ◽  
Author(s):  
Aparna Bansal ◽  
Himanshu
Keyword(s):  
Gene Therapy ◽  
Viral Vectors ◽  
Transfection Efficiency ◽  
Gene Transfection ◽  
Expression System ◽  
Target Cells ◽  
Specific Expression ◽  
Naked Dna

Introduction: Gene therapy has emerged out as a promising therapeutic pave for the treatment of genetic and acquired diseases. Gene transfection into target cells using naked DNA is a simple and safe approach which has been further improved by combining vectors or gene carriers. Both viral and non-viral approaches have achieved a milestone to establish this technique, but non-viral approaches have attained a significant attention because of their favourable properties like less immunotoxicity and biosafety, easy to produce with versatile surface modifications, etc. Literature is rich in evidences which revealed that undoubtedly, non–viral vectors have acquired a unique place in gene therapy but still there are number of challenges which are to be overcome to increase their effectiveness and prove them ideal gene vectors. Conclusion: To date, tissue specific expression, long lasting gene expression system, enhanced gene transfection efficiency has been achieved with improvement in delivery methods using non-viral vectors. This review mainly summarizes the various physical and chemical methods for gene transfer in vitro and in vivo.

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