scholarly journals A Reverse-Genetics Mutational Analysis of the Barley HvDWARF Gene Results in Identification of a Series of Alleles and Mutants with Short Stature of Various Degree and Disturbance in BR Biosynthesis Allowing a New Insight into the Process

2016 ◽  
Vol 17 (4) ◽  
pp. 600 ◽  
Author(s):  
Damian Gruszka ◽  
Malgorzata Gorniak ◽  
Ewelina Glodowska ◽  
Ewa Wierus ◽  
Jana Oklestkova ◽  
...  
Keyword(s):  
Short Stature ◽  
Reverse Genetics ◽  
Mutational Analysis ◽  
Insight Into

Mutational Analysis Provides Molecular Insight into the Carbohydrate-Binding Region of Calreticulin:  Pivotal Roles of Tyrosine-109 and Aspartate-135 in Carbohydrate Recognition†

Biochemistry ◽  
2004 ◽  
Vol 43 (1) ◽  
pp. 97-106 ◽  
Author(s):  
Mili Kapoor ◽  
Lars Ellgaard ◽  
Jayashree Gopalakrishnapai ◽  
Christiane Schirra ◽  
Emiliano Gemma ◽  
...  
Keyword(s):  
Mutational Analysis ◽  
Carbohydrate Binding ◽  
Binding Region ◽  
Carbohydrate Recognition ◽  
Insight Into

scholarly journals Optimizing RNA interference for application in mammalian cells

2004 ◽  
Vol 380 (3) ◽  
pp. 593-603 ◽  
Author(s):  
René H. MEDEMA
Keyword(s):  
Rna Interference ◽  
Gene Silencing ◽  
Mammalian Cells ◽  
Reverse Genetics ◽  
Mutational Analysis ◽  
Life Sciences ◽  
Novel Technologies ◽  
Genome Wide ◽  
Technological Developments ◽  
Interfering Rna

Over the last 2 years, the scientific community has rapidly embraced novel technologies that allow gene silencing in vertebrates. Ease of application, cost effectiveness and the possibilities for genome-wide reverse genetics have quickly turned this approach into a widely accepted, almost mandatory asset for a self-respecting laboratory in life sciences. This review discusses some of the recent technological developments that allow the application of RNAi (RNA interference) in mammalian cells. In addition, the advantages of applying RNAi to study cell cycle events and the emerging approaches to perform mutational analysis by complementation in mammalian cells are evaluated. In addition, common pitfalls and drawbacks of RNAi will be reviewed, as well as the possible ways to get around these shortcomings of gene silencing by small interfering RNA.

scholarly journals pFiD188, the Linear Virulence Plasmid of Rhodococcus fascians D188

2012 ◽  
Vol 25 (5) ◽  
pp. 637-647 ◽  
Author(s):  
Isolde Francis ◽  
Annick De Keyser ◽  
Philippe De Backer ◽  
Carmen Simón-Mateo ◽  
Jutta Kalkus ◽  
...  
Keyword(s):  
Virulence Genes ◽  
Mutational Analysis ◽  
Linear Plasmid ◽  
Virulence Plasmid ◽  
Symptom Development ◽  
Linear Plasmids ◽  
Type Iv ◽  
Rhodococcus Fascians ◽  
Syntenic Regions ◽  
Insight Into

Rhodococcus fascians is currently the only phytopathogen of which the virulence genes occur on a linear plasmid. To get insight into the origin of this replicon and into the virulence strategy of this broad-spectrum phytopathogen, the sequence of the linear plasmid of strain D188, pFiD188, was determined. Analysis of the 198,917 bp revealed four syntenic regions with linear plasmids of R. erythropolis, R. jostii, and R. opacus, suggesting a common origin of these replicons. Mutational analysis of pFi_086 and pFi_102, similar to cutinases and type IV peptidases, respectively, showed that conserved region R2 was involved in plasmid dispersal and pointed toward a novel function for actinobacterial cutinases in conjugation. Additionally, pFiD188 had three regions that were unique for R. fascians. Functional analysis of the stk and nrp loci of regions U2 and U3, respectively, indicated that their role in symptom development was limited compared with that of the previously identified fas, att, and hyp virulence loci situated in region U1. Thus, pFiD188 is a typical rhodococcal linear plasmid with a composite structure that encodes core functions involved in plasmid maintenance and accessory functions, some possibly acquired through horizontal gene transfer, implicated in virulence and the interaction with the host.

Mutational analysis of the CG recognizing DNA methyltransferase SssI: Insight into enzyme–DNA interactions

2009 ◽  
Vol 1794 (11) ◽  
pp. 1654-1662 ◽  
Author(s):  
Maria V. Darii ◽  
Natalia A. Cherepanova ◽  
Oksana M. Subach ◽  
Olga V. Kirsanova ◽  
Tamás Raskó ◽  
...  
Keyword(s):  
Dna Methyltransferase ◽  
Mutational Analysis ◽  
Dna Interactions ◽  
Insight Into

Mutational analysis of ProTx-I and the novel venom peptide Pe1b provide insight into residues responsible for selective inhibition of the analgesic drug target NaV1.7

2020 ◽  
Vol 181 ◽  
pp. 114080 ◽  
Author(s):  
Darshani B. Rupasinghe ◽  
Volker Herzig ◽  
Irina Vetter ◽  
Zoltan Dekan ◽  
John Gilchrist ◽  
...  
Keyword(s):  
Drug Target ◽  
Mutational Analysis ◽  
Selective Inhibition ◽  
The Novel ◽  
Analgesic Drug ◽  
Venom Peptide ◽  
Insight Into

scholarly journals Role of N-Linked Glycans on Bunyamwera Virus Glycoproteins in Intracellular Trafficking, Protein Folding, and Virus Infectivity

2005 ◽  
Vol 79 (21) ◽  
pp. 13725-13734 ◽  
Author(s):  
Xiaohong Shi ◽  
Kristina Brauburger ◽  
Richard M. Elliott
Keyword(s):  
Protein Folding ◽  
Intracellular Trafficking ◽  
Protein Misfolding ◽  
Reverse Genetics ◽  
Mutational Analysis ◽  
Glycosylation Site ◽  
Virus Infectivity ◽  
Membrane Glycoproteins ◽  
Recombinant Viruses ◽  
Bunyamwera Virus

ABSTRACT The membrane glycoproteins (Gn and Gc) of Bunyamwera virus (BUN, family Bunyaviridae) contain three potential sites for the attachment of N-linked glycans: one site (N60) on Gn and two (N624 and N1169) on Gc. We determined that all three sites are glycosylated. Digestion of the glycoproteins with endo-β-N-acetylglucosaminidase H (endo H) or peptide:N-glycosidase F revealed that Gn and Gc differ significantly in their glycan status and that late in infection Gc glycans remain endo H sensitive. The roles of the N-glycans in intracellular trafficking of the glycoproteins to the Golgi, protein folding, and virus replication were investigated by mutational analysis and confocal immunofluorescence. Elimination of the glycan on Gn, by changing N60 to a Q residue, resulted in the protein misfolding and failure of both Gn and Gc proteins to traffic to the Golgi complex. We were unable to rescue a viable virus by reverse genetics from a cDNA containing the N60Q mutation. In contrast, mutant Gc proteins lacking glycans on either N624 or N1169, or both sites, were able to target to the Golgi. Gc proteins containing mutations N624Q and N1169Q acquired endo H resistance. Three viable N glycosylation-site-deficient viruses, lacking glycans on one site or both sites on Gc, were created by reverse genetics. The viability of these recombinant viruses and analysis of growth kinetics indicates that the glycans on Gc are not essential for BUN replication, but they do contribute to the efficiency of virus infection.

scholarly journals Mutational Analysis of the vacA Promoter Provides Insight into Gene Transcription in Helicobacter pylori

1999 ◽  
Vol 181 (7) ◽  
pp. 2261-2266 ◽  
Author(s):  
Mark H. Forsyth ◽  
Timothy L. Cover
Keyword(s):  
Helicobacter Pylori ◽  
Gene Transcription ◽  
Functional Role ◽  
Mutational Analysis ◽  
Consensus Sequence ◽  
H Pylori ◽  
Insight Into

ABSTRACT Analysis of 12 Helicobacter pylori promoters indicates the existence of a consensus −10 hexamer (TAtaaT) but little conservation of −35 sequences. In this study, mutations in either the H. pylori vacA −10 region or the −35 region resulted in decreased vacA transcription and suggested that an extended −10 motif is utilized. Thus, despite the lack of a −35 consensus sequence for H. pylori promoters, the −35 region plays a functional role in vacA transcription.

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