Barley aleurone layer cell protoplasts as a transient expression system

1991 ◽  
Vol 16 (3) ◽  
pp. 463-467 ◽  
Author(s):  
Bhuvana Gopalakrishnan ◽  
Burachai Sonthayanon ◽  
Rownak Rahmatullah ◽  
Subbaratnam Muthukrishnan
Keyword(s):  
Transient Expression ◽  
Expression System ◽  
Aleurone Layer ◽  
Layer Cell ◽  
Transient Expression System ◽  
Barley Aleurone Layer ◽  
Barley Aleurone

scholarly journals Agroinfiltration of Strawberry Fruit — A Powerful Transient Expression System for Gene Validation

2016 ◽  
Vol 6 ◽  
pp. 19-37 ◽  
Author(s):  
Raquel F. Carvalho ◽  
Sofia D. Carvalho ◽  
Kevin O’Grady ◽  
Kevin M. Folta
Keyword(s):  
Transient Expression ◽  
Expression System ◽  
Strawberry Fruit ◽  
Transient Expression System

scholarly journals A Transient Expression System with Stably Integrated CRISPR-dCas9 Fusions for Regulation of Genes Involved in Immunoglobulin G Glycosylation

2022 ◽  
Author(s):  
Anika Mijakovac ◽  
Karlo Miškec ◽  
Jasminka Krištić ◽  
Vedrana Vičić Bočkor ◽  
Vanja Tadić ◽  
...  
Keyword(s):  
Transient Expression ◽  
Immunoglobulin G ◽  
Expression System ◽  
Transient Expression System

scholarly journals Functional characterization of a glycine 185-to-valine substitution in human P-glycoprotein by using a vaccinia-based transient expression system.

1996 ◽  
Vol 7 (10) ◽  
pp. 1485-1498 ◽  
Author(s):  
M Ramachandra ◽  
S V Ambudkar ◽  
M M Gottesman ◽  
I Pastan ◽  
C A Hrycyna
Keyword(s):  
Drug Resistance ◽  
Atpase Activity ◽  
Cyclosporin A ◽  
Transient Expression ◽  
Expression System ◽  
Rhodamine 123 ◽  
Wild Type ◽  
P Glycoprotein ◽  
Transient Expression System

Human P-glycoprotein (Pgp) is a 170-kDa plasma membrane protein that confers multidrug resistance to otherwise sensitive cells. A mutation in Pgp, G185-->V, originally identified as a spontaneous mutation, was shown previously to alter the drug resistance profiles in cell lines that are stably transfected with the mutant MDR1 cDNA and selected with cytotoxic agents. To understand the mechanism by which the V185 mutation leads to an altered drug resistance profile, we used a transient expression system that eliminates the need for drug selection to attain high expression levels and allows for the rapid characterization of many aspects of Pgp function and biosynthesis. The mutant and wild-type proteins were expressed at similar levels after 24-48 h in human osteosarcoma (HOS) cells by infection with a recombinant vaccinia virus encoding T7 RNA polymerase and simultaneous transfection with a plasmid containing MDR1 cDNA controlled by the T7 promoter. For both mutant and wild-type proteins, photolabeling with [3H]azidopine and [125I]iodoarylazidoprazosin, drug-stimulated ATPase activity, efflux of rhodamine 123, and accumulation of radiolabeled vinblastine and colchicine were evaluated. In crude membrane preparations from HOS cells, a higher level of basal Pgp-ATPase activity was observed for the V185 variant than for the wild-type, suggesting partial uncoupling of drug-dependent ATP hydrolysis by the mutant. Several compounds, including verapamil, nicardipine, tetraphenylphosphonium, and prazosin, stimulated ATPase activities of both the wild-type and mutant similarly, whereas cyclosporin A inhibited the ATPase activity of the mutant more efficiently than that of the wild-type. This latter observation explains the enhanced potency of cyclosporin A as an inhibitor of the mutant Pgp. No differences were seen in verapamil-inhibited rhodamine 123 efflux, but the rate of accumulation was slower for colchicine and faster for vinblastine in cells expressing the mutant protein, as compared with those expressing wild-type Pgp. We conclude that the G185-->V mutation confers pleiotropic alterations on Pgp, including an altered basal ATPase activity and altered interaction with substrates and the inhibitor cyclosporin A.

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