In vitroTranslation of Natural mRNAs in a Cell-free System Containing Components from Interferon-treated Chicken Fibroblasts and Factor Preparations from Mouse Ascites Cells or Rabbit Reticulocytes

1975 ◽  
Vol 30 (5-6) ◽  
pp. 398-405 ◽  
Author(s):  
G. Hiller ◽  
G. Viehhauser ◽  
I. Winkler ◽  
D. Pohl ◽  
C. Jungwirth ◽  
...  
Keyword(s):  
Chick Embryo ◽  
In Vitro Translation ◽  
Interferon Treatment ◽  
Mixed Cell ◽  
Free System ◽  
Free Protein ◽  
Cell Free System ◽  
Mouse Ascites ◽  
Rabbit Reticulocytes

Abstract Interferon Mechanism, in vitro Translation, Viral and Cellular mRNA, Chick Embryo Fibroblasts, Vaccinia Infection The effect of interferon has been studied in a mixed cell-free protein synthesizing system. Hemo­ globin (Hb) and Encephalomyocarditis virus (EMC) -RNA can be efficiently translated in vitro in a system containing S-30 lysates or run-off ribosomes from primary chick embryo fibroblasts (CEF) and a postmicrosomal supernatant from mouse ascites cells or a ribosomal-wash preparation from rabbit reticulocytes. Ribosomes prepared from CEF pretreated with high doses of homologous inter­ feron (500 units/ml) were able to translate Hb-RNA in the presence of heterologous factors with the same efficiency as ribosomes prepared from control cells. Translation of EMC-RNA was slightly reduced if ribosomes from interferon-treated cells were used in the mixed cell-free system, confirming previous reports. No inhibitory effect caused by interferon treatment of CEF cells could be detected on in vitro translation of natural mRNAs if the cells had, in addition to interferon treatment, been infected with vaccinia virus. Possible reasons for the different observations made with our cell-free protein synthesizing system from CEF and with cell-free systems prepared from mouse cells are discussed.

scholarly journals Posttranslational incorporation of contractile proteins into myofibrils in a cell-free system.

1988 ◽  
Vol 107 (2) ◽  
pp. 587-596 ◽  
Author(s):  
M Bouché ◽  
S M Goldfine ◽  
D A Fischman
Keyword(s):  
In Vitro Translation ◽  
Homologous Proteins ◽  
Free System ◽  
Cell Free System ◽  
Reticulocyte Lysate ◽  
Thin Filament Proteins ◽  
A Cell ◽  
The Many ◽  
Kinetic Equilibrium

The incorporation of newly synthesized protein into myofibrils has been examined in a cell-free system. Myofibrils were added to a reticulocyte lysate after the in vitro translation of muscle-specific poly(A)+RNA. Only a small number of the many synthesized proteins were found to associate with the exogenously added myofibrils. These proteins were all identified as sarcomeric components and had subunit mobilities (Mr) of 200, 140, 95, 86, 43, 38, 35, 25, 23, 20, and 18 kD. The association was rapid (t1/2 less than 15 min) and, for most of the proteins, relatively temperature insensitive. Except for a 43-kD polypeptide, tentatively identified as beta-actin, none of the proteins encoded by brain poly(A)+RNA associated with the myofibrils. When filaments made from purified myosin or actin were used as the "capture" substrates, only thick or thin filament proteins, respectively, were incorporated. Incorporation was substantially reduced when cross-linked myosin filaments were used. These results are compatible with a model in which proteins of the sarcomere are in kinetic equilibrium with homologous proteins in a soluble pool.

In vitro translation in a hamster brain cell-free system

1991 ◽  
Vol 37 (3) ◽  
pp. 191-198
Author(s):  
Georges R. Thériaul ◽  
Didier Gauthier
Keyword(s):  
Brain Cell ◽  
In Vitro Translation ◽  
Free System ◽  
Cell Free System

Biotin in vitro Translation: A Nonradioactive Method for the Synthesis of Biotin Labeled Proteins in a Cell-Free System

1996 ◽  
pp. 183-199
Author(s):  
Hans-Joachim Hoeltke ◽  
Irene Ettl ◽  
Edith Strobel ◽  
Hermann Leying ◽  
Maria Zimmermann ◽  
...  
Keyword(s):  
In Vitro Translation ◽  
Free System ◽  
Cell Free System ◽  
A Cell

scholarly journals Endogenous read-through of a UGA termination codon in a Saccharomyces cerevisiae cell-free system: evidence for involvement of both a mitochondrial and a nuclear tRNA.

1982 ◽  
Vol 2 (5) ◽  
pp. 490-497 ◽  
Author(s):  
M F Tuite ◽  
C S McLaughlin
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Deae Cellulose ◽  
In Vitro Translation ◽  
Translation System ◽  
Beta Globin ◽  
Mitochondrial Trna ◽  
Globin Mrna ◽  
Free System ◽  
Cell Free System

Globin mRNA, translated in a Saccharomyces cerevisiae cell-free protein synthesizing system prepared from a [psi+ rho+] strain, primarily directed the synthesis of alpha- and beta-globin. A third globin mRNA-specific polypeptide was also synthesized, representing approximately 10% of the total translation products. This polypeptide (beta') was synthesized by translational read-through of the beta- globin mRNA UGA terminator and was mediated primarily by an endogenous tRNA coded for by the mitochondria. This mitochondrial tRNA, when charged, could be preferentially bound, in high salt, to benzoylated DEAE-cellulose, a characteristic of a tRNATrp. The synthesis of beta- mediated by this mitochondrial tRNATrp was significantly reduced when the translation system was prepared from an isogenic [psi-] strain. Evidence for a nuclear-coded tRNA, also able to suppress the beta-globin mRNA UGA terminator in [psi+] but not [psi-] lysates, was also obtained. The presence of these endogenous UGA suppressor activities in the yeast cell-free system should allow successful in vitro translation of mitochondrial mRNAs.

In Vitro Translation in an Insect-Based Cell-Free System

2003 ◽  
pp. 209-217 ◽  
Author(s):  
Stefan Kubick ◽  
J. Schacherl ◽  
H. Fleischer-Notter ◽  
E. Royall ◽  
L. O. Roberts ◽  
...  
Keyword(s):  
In Vitro Translation ◽  
Free System ◽  
Cell Free System
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