Coupled in vitro transcription/translation based on wheat germ extract for efficient expression from PCR-generated templates in short-time batch reactions

2021 ◽  
pp. 128412
Author(s):  
Hajime Takahashi ◽  
Atsushi Ogawa
Keyword(s):  
Wheat Germ ◽  
In Vitro Transcription ◽  
Wheat Germ Extract ◽  
Efficient Expression ◽  
Short Time

scholarly journals Decrease in the Rate of Protein Synthesis by Polysomes from Cultured Fibroblasts of Patients and Carriers with Duchenne Muscular Dystrophy

1979 ◽  
Vol 6 (3) ◽  
pp. 355-358 ◽  
Author(s):  
M. Boulé ◽  
M. Vanasse ◽  
L. Brakier-Gingras
Keyword(s):  
Protein Synthesis ◽  
Duchenne Muscular Dystrophy ◽  
Muscular Dystrophy ◽  
Wheat Germ ◽  
Fibroblast Cells ◽  
Wheat Germ Extract ◽  
Cultured Fibroblasts ◽  
Normal Controls

SUMMARY:Polysomes extracted from cultured fibroblast cells isolated from patients with Duchenne muscular dystrophy (DMD), carriers of the disease, and normal controls were used for in vitro measurement of protein synthesis in a wheat germ extract system. It was observed that polysomes from patients and carriers (seven of each aged 17 years or older) exhibited a 3-fold and a 1.5-fold decrease in the rate of protein synthesis, respectively, as compared with controls. These results are discussed with a view to developing a sensitive and easily available assay for the detection of DMD carriers.

scholarly journals Construction of a synthetic messenger RNA encoding a membrane protein.

1983 ◽  
Vol 96 (5) ◽  
pp. 1464-1469 ◽  
Author(s):  
J L Rubenstein ◽  
T G Chappell
Keyword(s):  
G Protein ◽  
Messenger Rna ◽  
In Vitro Transcription ◽  
Membrane Insertion ◽  
Wheat Germ Extract ◽  
E Coli ◽  
Infected Cells ◽  
Vesicular Stomatitis ◽  
Synthetic Mrna

We have synthesized microgram quantities of a functional eucaryotic mRNA by in vitro transcription. For this purpose, we constructed a plasmid in which the Escherichia coli lactose promoter was 5' to the vesicular stomatitis virus (VSV) G protein gene (Rose, J. K., and C. J. Gallione, 1981, J. Virol., 39:519-528). This DNA served as the template in an in vitro transcription reaction utilizing E. coli RNA polymerase. The RNA product was capped using the vaccinia guanylyltransferase. A typical preparation of the synthetic G mRNA was equivalent to the amount of G mRNA that can be isolated from approximately 10(8) VSV-infected cells. This synthetic mRNA was translated by a wheat germ extract in the presence of microsomes, producing a polypeptide that was indistinguishable from G protein in its size, antigenicity, degree of glycosylation, and its membrane insertion. This technique should aid in identifying features needed by proteins for insertion into membranes.

Investigation of end processing and degradation of premature tRNAs and their application to stabilization of in vitro transcripts in wheat germ extract

2015 ◽  
Vol 13 (4) ◽  
pp. 1008-1012 ◽  
Author(s):  
Atsushi Ogawa ◽  
Yasunori Doi
Keyword(s):  
Wheat Germ ◽  
Wheat Germ Extract ◽  
In Vitro Transcripts

We investigated the end processing and degradation of premature tRNAs in wheat germ extract (left), which led to the findings of end protectors for efficiently stabilizing an in vitro transcript (purple, right).

Sign in / Sign up

Export Citation Format

Share Document