scholarly journals Establishing RNA Interference as a Reverse-Genetic Approach for Gene Functional Analysis in Protoplasts

2008 ◽  
Vol 149 (2) ◽  
pp. 642-652 ◽  
Author(s):  
Zhiyang Zhai ◽  
Thanwalee Sooksa-nguan ◽  
Olena K. Vatamaniuk
Keyword(s):  
Functional Analysis ◽  
Rna Interference ◽  
Reverse Genetic ◽  
Genetic Approach ◽  
Gene Functional Analysis ◽  
Reverse Genetic Approach

scholarly journals RNA-binding proteins and circadian rhythms in Arabidopsis thaliana

2001 ◽  
Vol 356 (1415) ◽  
pp. 1755-1759 ◽  
Author(s):  
Dorothee Staiger
Keyword(s):  
Arabidopsis Thaliana ◽  
Feedback Loop ◽  
Rna Binding ◽  
Rna Binding Proteins ◽  
Transcriptional Level ◽  
Reverse Genetic ◽  
Genetic Approach ◽  
Circadian Oscillators ◽  
Clock Proteins ◽  
Reverse Genetic Approach

An Arabidopsis transcript preferentially expressed at the end of the daily light period codes for the RNA–binding protein At GRP7. A reverse genetic approach in Arabidopsis thaliana has revealed its role in the generation of circadian rhythmicity: At GRP7 is part of a negative feedback loop through which it influences the oscillations of its own transcript. Biochemical and genetic experiments indicate a mechanism for this autoregulatory circuit: At grp7 gene transcription is rhythmically activated by the circadian clock during the day. The At GPR7 protein accumulates with a certain delay and represses further accumulation of its transcript, presumably at the post–transcriptional level. In this respect, the At GRP7 feedback loop differs from known circadian oscillators in the fruitfly Drosophila and mammals based on oscillating clock proteins that repress transcription of their own genes with a 24 h rhythm. It is proposed that the At GRP7 feedback loop may act within an output pathway from the Arabidopsis clock.

scholarly journals Structural and phenotypic analysis of Chikungunya virus RNA replication elements

2019 ◽  
Vol 47 (17) ◽  
pp. 9296-9312 ◽  
Author(s):  
Catherine Kendall ◽  
Henna Khalid ◽  
Marietta Müller ◽  
Dominic H Banda ◽  
Alain Kohl ◽  
...  
Keyword(s):  
Rational Design ◽  
Chikungunya Virus ◽  
Rna Replication ◽  
Host Cells ◽  
Genome Replication ◽  
Reverse Genetic ◽  
Genetic Approach ◽  
Phenotypic Analysis ◽  
Efficient Replication ◽  
Reverse Genetic Approach

Abstract Chikungunya virus (CHIKV) is a re-emerging, pathogenic Alphavirus transmitted to humans by Aedes spp. mosquitoes. We have mapped the RNA structure of the 5′ region of the CHIKV genome using selective 2′-hydroxyl acylation analysed by primer extension (SHAPE) to investigate intramolecular base-pairing at single-nucleotide resolution. Taking a structure-led reverse genetic approach, in both infectious virus and sub-genomic replicon systems, we identified six RNA replication elements essential to efficient CHIKV genome replication - including novel elements, either not previously analysed in other alphaviruses or specific to CHIKV. Importantly, through a reverse genetic approach we demonstrate that the replication elements function within the positive-strand genomic copy of the virus genome, in predominantly structure-dependent mechanisms during efficient replication of the CHIKV genome. Comparative analysis in human and mosquito-derived cell lines reveal that a novel element within the 5′UTR is essential for efficient replication in both host systems, while those in the adjacent nsP1 encoding region are specific to either vertebrate or invertebrate host cells. In addition to furthering our knowledge of fundamental aspects of the molecular virology of this important human pathogen, we foresee that results from this study will be important for rational design of a genetically stable attenuated vaccine.

scholarly journals A reverse genetic approach for generating gene replacement mutants in Ustilago maydis

2004 ◽  
Vol 272 (4) ◽  
pp. 488-488 ◽  
Author(s):  
A. Brachmann ◽  
J. König ◽  
C. Julius ◽  
M. Feldbrügge
Keyword(s):  
Ustilago Maydis ◽  
Gene Replacement ◽  
Reverse Genetic ◽  
Genetic Approach ◽  
Reverse Genetic Approach

Aminopeptidases and dipeptidyl-peptidases secreted by the dermatophyte Trichophyton rubrum

Microbiology ◽  
2005 ◽  
Vol 151 (1) ◽  
pp. 145-155 ◽  
Author(s):  
Michel Monod ◽  
Barbara Léchenne ◽  
Olivier Jousson ◽  
Daniela Grand ◽  
Christophe Zaugg ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Trichophyton Rubrum ◽  
Hydrolytic Activity ◽  
Cdna Libraries ◽  
Sole Nitrogen Source ◽  
Reverse Genetic ◽  
Genetic Approach ◽  
Leucine Aminopeptidases ◽  
Dipeptidyl Peptidases ◽  
Reverse Genetic Approach

The nature of secreted aminopeptidases in Trichophyton rubrum was investigated by using a reverse genetic approach. T. rubrum genomic and cDNA libraries were screened with Aspergillus spp. and Saccharomyces cerevisiae aminopeptidase genes as the probes. Two leucine aminopeptidases, ruLap1 and ruLap2, and two dipeptidyl-peptidases, ruDppIV and ruDppV, were characterized and compared to orthologues secreted by Aspergillus fumigatus using a recombinant protein from Pichia pastoris. RuLap1 is a 33 kDa nonglycosylated protein, while ruLap2 is a 58–65 kDa glycoprotein. The hydrolytic activity of ruLap1, ruLap2 and A. fumigatus orthologues showed various preferences for different aminoacyl-7-amido-4-methylcoumarin substrates, and various sensitivities to inhibitors and cations. ruDppIV and ruDppV showed similar activities to A. fumigatus orthologues. In addition to endopeptidases, the four aminopeptidases ruLap1, ruLap2, ruDppIV and ruDppV were produced by T. rubrum in a medium containing keratin as the sole nitrogen source. Synergism between endo- and exopeptidases is likely to be essential for dermatophyte virulence, since these fungi grow only in keratinized tissues.

A reverse genetic approach for generating gene replacement mutants in Ustilago maydis

2004 ◽  
Vol 272 (2) ◽  
pp. 216-226 ◽  
Author(s):  
A. Brachmann ◽  
J. König ◽  
C. Julius ◽  
M. Feldbrügge
Keyword(s):  
Ustilago Maydis ◽  
Gene Replacement ◽  
Reverse Genetic ◽  
Genetic Approach ◽  
Reverse Genetic Approach

Marker reconstitution mutagenesis: a simple and efficient reverse genetic approach

Yeast ◽  
2010 ◽  
Vol 28 (3) ◽  
pp. 205-212 ◽  
Author(s):  
Xie Tang ◽  
Junqi Huang ◽  
Anup Padmanabhan ◽  
Kavya Bakka ◽  
Yun Bao ◽  
...  
Keyword(s):  
Reverse Genetic ◽  
Genetic Approach ◽  
Reverse Genetic Approach

A novel reverse-genetic approach (SIMF) identifies Mutator insertions in new Myb genes

Planta ◽  
2000 ◽  
Vol 211 (6) ◽  
pp. 887-893 ◽  
Author(s):  
Pablo D. Rabinowicz ◽  
Erich Grotewold
Keyword(s):  
Reverse Genetic ◽  
Genetic Approach ◽  
Myb Genes ◽  
Reverse Genetic Approach

scholarly journals Reverse Genetic Approach to Exploring Genes Responsible for Cell-Wall Dynamics in Supporting Tissues of Arabidopsis thaliana under Microgravity Conditions

2007 ◽  
Vol 21 (3) ◽  
pp. 48-55 ◽  
Author(s):  
Kento Koizumi ◽  
Ryusuke Yokoyama ◽  
Motoshi Kamada ◽  
Katsunori Omori ◽  
Noriaki Ishioka ◽  
...  
Keyword(s):  
Arabidopsis Thaliana ◽  
Cell Wall ◽  
Reverse Genetic ◽  
Genetic Approach ◽  
Microgravity Conditions ◽  
Reverse Genetic Approach
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