scholarly journals In Vivo Precision Genetic Change of Soybean Δ9-Stearoyl (18:0)-ACP Desaturase Gene for Increase Palmitoyl (16:0)-ACP Specificity and along with Change of Endogenous ALS Genes as Selectable Markers and Yeast Strain “EY 957” as a Model

2021 ◽  
Vol 05 (03) ◽  
pp. 223-237
Author(s):  
Lewamy Mamadou ◽  
Suryadevara S. Rao ◽  
David Hildebrand
Keyword(s):  
Yeast Strain ◽  
Genetic Change ◽  
Desaturase Gene ◽  
Selectable Markers

scholarly journals Shuttle Vector Expression in Thermococcus kodakaraensis: Contributions of cis Elements to Protein Synthesis in a Hyperthermophilic Archaeon

2008 ◽  
Vol 74 (10) ◽  
pp. 3099-3104 ◽  
Author(s):  
Thomas J. Santangelo ◽  
L'ubomíra Čuboňová ◽  
John N. Reeve
Keyword(s):  
Translation Initiation ◽  
Affinity Purification ◽  
Shuttle Vector ◽  
Selectable Markers ◽  
Hyperthermophilic Archaeon ◽  
Shuttle Vectors ◽  
Ribosome Binding ◽  
Translation Initiation Codon ◽  
Binding Sequence

ABSTRACT Shuttle vectors that replicate stably and express selectable phenotypes in both Thermococcus kodakaraensis and Escherichia coli have been constructed. Plasmid pTN1 from Thermococcus nautilis was ligated to the commercial vector pCR2.1-TOPO, and selectable markers were added so that T. kodakaraensis transformants could be selected by ΔtrpE complementation and/or mevinolin resistance. Based on Western blot measurements, shuttle vector expression of RpoL-HA, a hemagglutinin (HA) epitope-tagged subunit of T. kodakaraensis RNA polymerase (RNAP), was ∼8-fold higher than chromosome expression. An idealized ribosome binding sequence (5′-AGGTGG) was incorporated for RpoL-HA expression, and changes to this sequence reduced expression. Changing the translation initiation codon from AUG to GUG did not reduce RpoL-HA expression, but replacing AUG with UUG dramatically reduced RpoL-HA synthesis. When functioning as translation initiation codons, AUG, GUG, and UUG all directed the incorporation of methionine as the N-terminal residue of RpoL-HA synthesized in T. kodakaraensis. Affinity purification confirmed that an HA- plus six-histidine-tagged RpoL subunit (RpoL-HA-his6) synthesized ectopically from a shuttle vector was assembled in vivo into RNAP holoenzymes that were active and could be purified directly from T. kodakaraensis cell lysates by Ni2+ binding and imidazole elution.

scholarly journals Dihydroceramide desaturase directs the switch between exosome production and autophagy for neuronal maintenance

2020 ◽  
Author(s):  
Chen-Yi Wu ◽  
Jhih-Gang Jhang ◽  
Chih-Wei Lin ◽  
Han-Chen Ho ◽  
Chih-Chiang Chan ◽  
...  
Keyword(s):  
Nervous System ◽  
Desaturase Gene ◽  
Dihydroceramide Desaturase ◽  
Autophagic Degradation ◽  
Photoreceptor Neurons ◽  
And Function ◽  
Neuronal Maintenance ◽  
Autophagy Inhibition

ABSTRACTExosomes play important roles in the nervous system. Mutations in the human dihydroceramide desaturase gene, DEGS1, are recently linked to severe neurological disorders, but the cause remains unknown. Here, we show that Ifc is required for the morphology and function of Drosophila photoreceptor neurons and not in the surrounding glia, but the degeneration of ifc-KO eyes can be rescued by glial expression of ifc, possibly mediated by exosomes. We develop an in vivo assay using Drosophila eye imaginal discs and show that the level and activity of Ifc correlates with the detection of exosome-like vesicles. While ifc overexpression and autophagy inhibition both enhances exosome production, combining the two had no additive effect. Moreover, ifc-KO reduces the density of the exosome precursor intraluminal vesicles (ILVs) in vivo, and DEGS1 promotes ILV formation in vitro. In conclusion, dihydroceramide desaturase promotes exosome formation and prevents its autophagic degradation in the nervous system.

In vitro embryo production from bovine oocyte donors aspirated at different frequencies or following treatment with FSH

1996 ◽  
Vol 1996 ◽  
pp. 68-68
Author(s):  
K.L. Goodhand ◽  
R.G. Watt ◽  
M.E. Staines ◽  
L.C. Higgins ◽  
P.J. Broadbent ◽  
...  
Keyword(s):  
Transvaginal Ultrasound ◽  
Genetic Change ◽  
Bovine Oocyte ◽  
Ultrasound Guided ◽  
Embryo Production ◽  
Follicular Aspiration ◽  
Oocyte Donors ◽  
Pre Treatment

The combination of in vivo recovery of oocytes using transvaginal ultrasound guided aspiration and subsequent in vitro embryo production can be used to increase the rate of genetic change for efficiency of beef production by increasing selection intensity and reducing generation interval. The total number of oocytes recovered by aspiration and embryos produced is directly proportional to the number of aspiration sessions whether recovery takes place once or twice weekly. Pre-treatment of oocyte donors with FSH has been shown to improve the number of follicles available for aspiration but effects on embryo production have been conflicting (Bungartz et al., 1995; Goodhand et al., in press). The objective of this experiment was to compare the effect on embryo production of frequency of follicular aspiration and pre-treatment of donor cattle with FSH.

In vitro and in vivo manipulations of bacteriophage Mu DNA: Cloning of Mu ends and construction of mini-Mu's carrying selectable markers

Gene ◽  
1981 ◽  
Vol 13 (1) ◽  
pp. 37-46 ◽  
Author(s):  
George Chaconas ◽  
Frans J. de Bruijn ◽  
Malcolm J. Casadaban ◽  
James R. Lupski ◽  
T.Jesse Kwoh ◽  
...  
Keyword(s):  
Bacteriophage Mu ◽  
Selectable Markers ◽  
Dna Cloning

Rat hepatic vinyl chloride metabolites induce gene conversion in the yeast strain D7RAD in vitro and in vivo

1981 ◽  
Vol 91 (4-5) ◽  
pp. 381-390 ◽  
Author(s):  
F. Eckardt ◽  
H. Muliawan ◽  
N. de Ruiter ◽  
H. Kappus
Keyword(s):  
Gene Conversion ◽  
Vinyl Chloride ◽  
Yeast Strain

SsHKT1;1 is a potassium transporter of the C3 halophyte Suaeda salsa that is involved in salt tolerance

2014 ◽  
Vol 41 (8) ◽  
pp. 790 ◽  
Author(s):  
Qun Shao ◽  
Ning Han ◽  
Tonglou Ding ◽  
Feng Zhou ◽  
Baoshan Wang
Keyword(s):  
Salt Tolerance ◽  
Yeast Strain ◽  
Suaeda Salsa ◽  
K Uptake ◽  
Potassium Transporter ◽  
Cation Homeostasis ◽  
Heterologous Expression Systems ◽  
K Concentration ◽  
Low K

SsHKT1;1, a HKT1 homologue, was isolated from the C3 halophyte Suaeda salsa L. and its ion transport properties were investigated in heterologous systems. The expression of SsHKT1;1 suppressed a K+ transport-defective phenotype of the yeast strain CY162 (Δtrk1Δtrk2), suggesting the enhancement of K+ uptake with SsHKT1;1. However, it did not suppress the salt-sensitive phenotype of the yeast strain G19 (Δena1–4), which lacks a major component of Na+ efflux. Transgenic Arabidopsis thaliana (L.) Heynh. plants overexpressing SsHKT1;1 showed enhanced salt tolerance and increased shoot K+ concentration, whereas no significant changes in shoot Na+ concentration were observed. S. salsa was also used to investigate K+ uptake properties under salinity. The K+ transporters in the roots selectively mediated K+ uptake irrespective of external Na+ and their inhibitor did not affect Na+ uptake at low K+. Thus, both molecular and physiological studies provide strong in vivo evidence that SsHKT1;1 mainly acts as a potassium transporter in heterologous expression systems and S. salsa, and that it is involved in salt tolerance by taking part in the maintenance of cytosolic cation homeostasis, particularly, in the maintenance of K+ nutrition under salinity.

scholarly journals Expression of the maize MnSod (Sod3) gene in MnSOD-deficient yeast rescues the mutant yeast under oxidative stress.

Genetics ◽  
1992 ◽  
Vol 131 (4) ◽  
pp. 803-809 ◽  
Author(s):  
D Zhu ◽  
J G Scandalios
Keyword(s):  
Oxidative Stress ◽  
Yeast Strain ◽  
Functional Role ◽  
Transit Peptide ◽  
Yeast Cells ◽  
Mitochondrial Matrix ◽  
Higher Plant ◽  
Initial Portion

Abstract Superoxide dismutases (SOD) are ubiquitous in aerobic organisms and are believed to play a significant role in protecting cells against the toxic, often lethal, effect of oxygen free radicals. However, direct evidence that SOD does in fact participate in such a protective role is scant. The MnSOD-deficient yeast strain (Sod2d) offered an opportunity to test the functional role of one of several SOD isozymes from the higher plant maize in hopes of establishing a functional bioassay for other SODs. Herein, we present evidence that MnSOD functions to protect cells from oxidative stress and that this function is conserved between species. The maize Sod3 gene was introduced into the yeast strain Sod2d where it was properly expressed and its product processed into the yeast mitochondrial matrix and assembled into the functional homotetramer. Most significantly, expression of the maize Sod3 transgene in yeast rendered the transformed yeast cells resistant to paraquat-induced oxidative stress by complementing the MnSOD deficiency. Furthermore, analyses with various deletion mutants of the maize SOD-3 transit peptide in the MnSOD-deficient yeast strain indicate that the initial portion (about 8 amino acids) of the maize transit peptide is required to direct the protein into the yeast mitochondrial matrix in vivo to function properly. These findings indicate that the functional role of maize MnSOD is conserved and dependent on its proper subcellular location in the mitochondria of a heterologous system.

In vitro embryo production from bovine oocyte donors aspirated at different frequencies or following treatment with FSH

1996 ◽  
Vol 1996 ◽  
pp. 68-68
Author(s):  
K.L. Goodhand ◽  
R.G. Watt ◽  
M.E. Staines ◽  
L.C. Higgins ◽  
P.J. Broadbent ◽  
...  
Keyword(s):  
Transvaginal Ultrasound ◽  
Genetic Change ◽  
Bovine Oocyte ◽  
Ultrasound Guided ◽  
Embryo Production ◽  
Follicular Aspiration ◽  
Oocyte Donors ◽  
Pre Treatment

The combination of in vivo recovery of oocytes using transvaginal ultrasound guided aspiration and subsequent in vitro embryo production can be used to increase the rate of genetic change for efficiency of beef production by increasing selection intensity and reducing generation interval. The total number of oocytes recovered by aspiration and embryos produced is directly proportional to the number of aspiration sessions whether recovery takes place once or twice weekly. Pre-treatment of oocyte donors with FSH has been shown to improve the number of follicles available for aspiration but effects on embryo production have been conflicting (Bungartz et al., 1995; Goodhand et al., in press). The objective of this experiment was to compare the effect on embryo production of frequency of follicular aspiration and pre-treatment of donor cattle with FSH.

Beneficial effects of Saccharomyces cerevisiae RC016 in weaned piglets: in vivo and ex vivo analysis

2019 ◽  
Vol 10 (1) ◽  
pp. 33-42 ◽  
Author(s):  
G.R. Garcia ◽  
C.A. Dogi ◽  
V.L. Poloni ◽  
A.S. Fochesato ◽  
A. De Moreno de Leblanc ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Small Intestine ◽  
Yeast Strain ◽  
Intestinal Inflammation ◽  
Ex Vivo ◽  
Goblet Cells ◽  
Feed Additives ◽  
Weaned Piglets ◽  
Ex Vivo Model

Probiotics represents an alternative to replace antibiotics as growth promoters in animal feed and are able to control enteric bacterial diseases and to improve gut immunity. Saccharomyces cerevisiae RC016 showed previously inhibition/coagregation of pathogens) and mycotoxins adsorbent ability (aflatoxin B1, ochratoxin A and zearalenone). The aim of this work was to evaluate beneficial properties of S. cerevisiae RC016 in a non-inflammatory in vivo model in weaned piglets and in an intestinal inflammation ex vivo model induced by the mycotoxin deoxynivalenol (DON). Secretory immunoglobulin A (s-IgA) levels, intestinal cytokines, goblet cells and production parameters were evaluated in a pig model. For the in vivo assays, twelve pigs were weaned at 21 days and assigned to two groups: Control (n=6) and Yeast (n=6). Animals received yeast strain for three weeks. After 22 days the small intestine was recovered for determination of goblet cells and s-IgA. For the ex vivo assay, jejunal explants were obtained from 5 weeks old crossbred piglets and treated as follow: (1) control; (2) treated for 3 h with 10 μM DON used as an inflammatory stressor; (3) incubated with 107 cfu/ml yeast strain; (4) pre-incubated 1 h with 107 cfu/ml yeast strain and then treated for 3 h with 10 μM DON. CCL20, interleukin (IL)-1β, IL-8 and IL-22 gene expression was determined by qPCR. Oral administration of S. cerevisiae RC016 increased s-IgA, the number of goblet cells in small intestine and all the growth parameters measured. In the ex vivo model, the cytokine profile studied showed a potential anti-inflammatory effect of the administration of the yeast. In conclusion, S. cerevisiae RC016 is a promising candidate for feed additives formulation to improve animal growth and gut immune system. This yeast strain could be able to improve the gut health through counteracting the weaning-associated intestinal inflammation in piglets.

Sign in / Sign up

Export Citation Format

Share Document