scholarly journals MultiSite Gateway Technology Is Useful for Donor DNA Plasmid Construction in CRISPR/Cas9-Mediated Knock-In System

2019 ◽  
Author(s):  
Takeshi Yasuda ◽  
Katsushi Tajima
Keyword(s):  
Plasmid Construction ◽  
Dna Plasmid ◽  
Gateway Technology

scholarly journals Development of stable transgenic maize plants tolerant for drought by manipulating ABA signaling through Agrobacterium-mediated transformation

2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Sridevi Muppala ◽  
Pavan Kumar Gudlavalleti ◽  
Kodandarami Reddy Malireddy ◽  
Sateesh Kumar Puligundla ◽  
Premalatha Dasari
Keyword(s):  
Abiotic Stress ◽  
Inbred Line ◽  
Biochemical Characterization ◽  
Abiotic Stress Tolerance ◽  
Crop Improvement ◽  
Maximum Frequency ◽  
Aba Signaling ◽  
Tolerant Plants ◽  
Maize Plants ◽  
Gateway Technology

Abstract Background In crop plants, to cope up with the demand of food for rising population, revolutionary crop improvement programmes are being implemented for higher and higher yields. Abiotic stress, especially at flowering stage, causes drastic effect on yield in plants. Deforestation and urbanization made the water table very low and changed the climate which led to untimely and unforeseen rains which affect the yield of a crop through stress, both by lack of water as well as water logging (abiotic stress). Development of tolerant plants through breeding is a time-consuming programme and does not perform well in normal conditions. Development of stress-tolerant plants through transgenic technology is the better solution. Maize is a major crop used as food and fodder and has the commercial value in ethanol production. Hence, the genes viz., nced (9-cis-epoxycarotenoid dioxygenase) and rpk (receptor-like protein kinase), which play the key roles in the abscisic acid pathway and upstream component in ABA signaling have been transferred into maize plants through Agrobacterium-mediated transformation by optimizing several parameters to obtain maximum frequency of transformation. Results Cultures raised from immature embryos of 2-mm size isolated from maize cobs, 12–15 days after pollination, were used for transformation. rpk and nced genes under the control of leaP and salT promoters respectively, cloned using gateway technology, have been introduced into elite maize inbred lines. Maximum frequency of transformation was observed with the callus infected after 20 days of inoculation by using 100 μM acetosyringone, 10 min infection time, and 2 days incubation period after co-cultivation resulted in maximum frequency of transformation (6%) in the NM5884 inbred line. Integration of the genes has been confirmed with molecular characterization by performing PCRs with marker as well as gene-specific primers and through southern hybridization. Physiological and biochemical characterization was done in vitro (artificial stress) and in vivo (pot experiments). Conclusions Changes in the parameters which affect the transformation frequency yielded maximum frequency of transformation with 20-day-old callus in the NM5884 inbred line. Introducing two or more genes using gateway technology is useful for developing stable transgenic plants with desired characters, abiotic stress tolerance in this study.

scholarly journals Secretion of killer toxin encoded on the linear DNA plasmid pGKL1 from Saccharomyces cerevisiae.

1990 ◽  
Vol 265 (28) ◽  
pp. 17274-17280
Author(s):  
M Tokunaga ◽  
A Kawamura ◽  
K Kitada ◽  
F Hishinuma
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Killer Toxin ◽  
Linear Dna ◽  
Dna Plasmid

scholarly journals Optimization of a New Non-viral Vector for Transfection: Eudragit Nanoparticles for the Delivery of a DNA Plasmid

2009 ◽  
Vol 8 (6) ◽  
pp. 433-443 ◽  
Author(s):  
M. Gargouri ◽  
A. Sapin ◽  
S. Bouali ◽  
P. Becuwe ◽  
JL Merlin ◽  
...  
Keyword(s):  
Viral Vector ◽  
Dna Plasmid

scholarly journals Keratinocyte growth factor enhances DNA plasmid tumor vaccine responses after murine allogeneic bone marrow transplantation

Blood ◽  
2009 ◽  
Vol 113 (7) ◽  
pp. 1574-1580 ◽  
Author(s):  
Robert R. Jenq ◽  
Christopher G. King ◽  
Christine Volk ◽  
David Suh ◽  
Odette M. Smith ◽  
...  
Keyword(s):  
Bone Marrow ◽  
T Cells ◽  
T Cell ◽  
Keratinocyte Growth Factor ◽  
Allogeneic Bone Marrow Transplantation ◽  
Allogeneic Bone ◽  
Effector Cells ◽  
Cd8 Cells ◽  
T Cell Reconstitution ◽  
Dna Plasmid

Abstract Keratinocyte growth factor (KGF), which is given exogenously to allogeneic bone marrow transplantation (allo-BMT) recipients, supports thymic epithelial cells and increases thymic output of naive T cells. Here, we demonstrate that this improved T-cell reconstitution leads to enhanced responses to DNA plasmid tumor vaccination. Tumor-bearing mice treated with KGF and DNA vaccination have improved long-term survival and decreased tumor burden after allo-BMT. When assayed before vaccination, KGF-treated allo-BMT recipients have increased numbers of peripheral T cells, including CD8+ T cells with vaccine-recognition potential. In response to vaccination, KGF-treated allo-BMT recipients, compared with control subjects, generate increased numbers of tumor-specific CD8+ cells, as well as increased numbers of CD8+ cells producing interferon-γ (IFN-γ) and tumor necrosis factor-α (TNF-α). We also found unanticipated benefits to antitumor immunity with the administration of KGF. KGF-treated allo-BMT recipients have an improved ratio of T effector cells to regulatory T cells, a larger fraction of effector cells that display a central memory phenotype, and effector cells that are derived from a broader T-cell–receptor repertoire. In conclusion, our data suggest that KGF can function as a potent vaccine adjuvant after allo-BMT through its effects on posttransplantation T-cell reconstitution.

scholarly journals Protective efficacy of a prime-boost protocol using H5-DNA plasmid as prime and inactivated H5N2 vaccine as the booster against the Egyptian avian influenza challenge virus

2016 ◽  
Vol 60 (03) ◽  
pp. 307-315
Author(s):  
H. A. HUSSEIN ◽  
B. M. AHMED ◽  
S. M. ALY ◽  
A. H. EL-DEEB ◽  
A. A. EL-SANOUSI ◽  
...  
Keyword(s):  
Avian Influenza ◽  
Protective Efficacy ◽  
Challenge Virus ◽  
Dna Plasmid

scholarly journals Development of the Multiple Gene Knockout System with One-Step PCR in Thermoacidophilic Crenarchaeon Sulfolobus acidocaldarius

Archaea ◽  
2017 ◽  
Vol 2017 ◽  
pp. 1-12 ◽  
Author(s):  
Shoji Suzuki ◽  
Norio Kurosawa
Keyword(s):  
Gene Knockout ◽  
Arginine Decarboxylase ◽  
Sulfolobus Acidocaldarius ◽  
Dna Photolyase ◽  
Genetic Studies ◽  
Multiple Gene ◽  
Plasmid Construction ◽  
Pcr Product ◽  
Optimized Protocol ◽  
One Step

Multiple gene knockout systems developed in the thermoacidophilic crenarchaeon Sulfolobus acidocaldarius are powerful genetic tools. However, plasmid construction typically requires several steps. Alternatively, PCR tailing for high-throughput gene disruption was also developed in S. acidocaldarius, but repeated gene knockout based on PCR tailing has been limited due to lack of a genetic marker system. In this study, we demonstrated efficient homologous recombination frequency (2.8 × 104 ± 6.9 × 103 colonies/μg DNA) by optimizing the transformation conditions. This optimized protocol allowed to develop reliable gene knockout via double crossover using short homologous arms and to establish the multiple gene knockout system with one-step PCR (MONSTER). In the MONSTER, a multiple gene knockout cassette was simply and rapidly constructed by one-step PCR without plasmid construction, and the PCR product can be immediately used for target gene deletion. As an example of the applications of this strategy, we successfully made a DNA photolyase- (phr-) and arginine decarboxylase- (argD-) deficient strain of S. acidocaldarius. In addition, an agmatine selection system consisting of an agmatine-auxotrophic strain and argD marker was also established. The MONSTER provides an alternative strategy that enables the very simple construction of multiple gene knockout cassettes for genetic studies in S. acidocaldarius.

Development of Marine Diesel Engine Database Based on Web

2014 ◽  
Vol 556-562 ◽  
pp. 5280-5283 ◽  
Author(s):  
Shou Wen Pang ◽  
Xiu Fu Song
Keyword(s):  
Diesel Engine ◽  
System Architecture ◽  
System Development ◽  
Marine Diesel Engine ◽  
Web Database ◽  
Development Environment ◽  
Network Education ◽  
Multimedia Teaching ◽  
Marine Diesel ◽  
Gateway Technology

The combination of marine diesel engine traditional multimedia teaching material database and Web database developed the Web database based on. The system development environment, system function, system architecture and database gateway technology are analyzed in the paper. Further this database plays a role in the network education in now.

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