Wheat Germ Cell-Free Translation System as a Tool for In vitro Selection of Functional Proteins

2002 ◽  
Vol 5 (6) ◽  
pp. 473-480
Author(s):  
Bentham Science Publisher A.N. Alexandrov ◽  
Bentham Science Publisher V.Yu. Alakhov ◽  
Bentham Science Publisher A.I. Miroshnikov
Keyword(s):  
Germ Cell ◽  
Wheat Germ ◽  
In Vitro Selection ◽  
Translation System ◽  
Free Translation ◽  
Selection Of

scholarly journals Gene expression in Acetabularia. I. Calibration of wheat germ cell-free translation system proteins as internal references for two-dimensional electrophoresis

1982 ◽  
Vol 58 (1) ◽  
pp. 23-33
Author(s):  
R.L. Shoeman ◽  
H.G. Schweiger
Keyword(s):  
Germ Cell ◽  
Isoelectric Point ◽  
Wheat Germ ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
In Vitro Translation ◽  
Translation System ◽  
Two Dimensional ◽  
Free Translation

Modification of existing two-dimensional techniques enables isoelectric focusing and sodium dodecyl sulphate polyacrylamide gel electrophoresis of complex mixtures of proteins to be completed within 8 h. The method was optimized to separate the protein components of a wheat germ cell-free translation system, providing a statistically proven resolution better than 0. 03 of a pH unit for the isoelectric point and 1000 for Mr. Fourteen of the more than 300 proteins separated were characterized with respect to Mr and isoelectric point relative to standard proteins under the same conditions. Stained wheat germ proteins thus serve as internal standards for analysis of in vitro translation products.

Cell-free synthesis of rat liver zinc-thioneins

1979 ◽  
Vol 57 (7) ◽  
pp. 1030-1035 ◽  
Author(s):  
Choy L. Hew ◽  
Peter E. Penner
Keyword(s):  
Germ Cell ◽  
Rat Liver ◽  
Gel Electrophoresis ◽  
Zinc Sulfate ◽  
Trichloroacetic Acid ◽  
Wheat Germ ◽  
Translation System ◽  
Glycine Buffer ◽  
Free Translation

The induction of rat liver zinc-thioneins mRNA was studied in a wheat germ cell-free translation system. Liver poly A rich polysomal RNA was isolated from rats which had been injected with zinc sulfate 5 h previously. These RNA preparations stimulated the incorporation of [35S]cystine into trichloroacetic acid insoluble proteins when assayed in the cell-free synthetic system. The translation products were characterized by Sephadex G-75 chromatography in 8 M urea – 50 mM β-mercaptoethanol, by disc gel electrophoresis in 4 M urea – Tris–glycine buffer (pH 9.2), and by peptide fingerprinting with pepsin. These results were identical with authentic rat liver zinc-thioneins. The zinc-thioneins mRNA activity in the control rats, however, was minimal. The stimulation in zinc-thioneins synthesis observed in the cell-free synthesis was similar to the increased synthesis of these polypeptides in vivo.

Production of yeast tRNA (m7G46) methyltransferase (Trm8–Trm82 complex) in a wheat germ cell-free translation system

2008 ◽  
Vol 133 (4) ◽  
pp. 453-460 ◽  
Author(s):  
Keisuke Matsumoto ◽  
Chie Tomikawa ◽  
Takashi Toyooka ◽  
Anna Ochi ◽  
Yoshitaka Takano ◽  
...  
Keyword(s):  
Germ Cell ◽  
Wheat Germ ◽  
Translation System ◽  
Yeast Trna ◽  
Free Translation

scholarly journals Hetero subunit interaction and RNA recognition of yeast tRNA (m7G46) methyltransferase synthesized in a wheat germ cell-free translation system

2007 ◽  
Vol 51 (1) ◽  
pp. 359-360 ◽  
Author(s):  
Y. Muneyoshi ◽  
K. Matsumoto ◽  
C. Tomikawa ◽  
T. Toyooka ◽  
A. Ochi ◽  
...  
Keyword(s):  
Germ Cell ◽  
Wheat Germ ◽  
Translation System ◽  
Rna Recognition ◽  
Subunit Interaction ◽  
Yeast Trna ◽  
Free Translation

scholarly journals The consensus motif for N-myristoylation of plant proteins in a wheat germ cell-free translation system

FEBS Journal ◽  
2010 ◽  
Vol 277 (17) ◽  
pp. 3596-3607 ◽  
Author(s):  
Seiji Yamauchi ◽  
Naoki Fusada ◽  
Hidenori Hayashi ◽  
Toshihiko Utsumi ◽  
Nobuyuki Uozumi ◽  
...  
Keyword(s):  
Germ Cell ◽  
Wheat Germ ◽  
Translation System ◽  
Plant Proteins ◽  
Consensus Motif ◽  
Free Translation

scholarly journals A Concept for Selection of Codon-Suppressor tRNAs Based on Read-Through Ribosome Display in anIn VitroCompartmentalized Cell-Free Translation System

2012 ◽  
Vol 2012 ◽  
pp. 1-7 ◽  
Author(s):  
Atsushi Ogawa ◽  
Masayoshi Hayami ◽  
Shinsuke Sando ◽  
Yasuhiro Aoyama
Keyword(s):  
Stop Codon ◽  
Full Length ◽  
Translation System ◽  
Anticodon Loop ◽  
Ribosome Display ◽  
Amber Stop Codon ◽  
Free Translation ◽  
Translatable Mrna ◽  
Selection Of

Here is presented a concept forin vitroselection of suppressor tRNAs. It uses a pool of dsDNA templates in compartmentalized water-in-oil micelles. The template contains a transcription/translation trigger, an amber stop codon, and another transcription trigger for the anticodon- or anticodon loop-randomized gene for tRNASer. Upon transcription are generated two types of RNAs, a tRNA and a translatable mRNA (mRNA-tRNA). When the tRNA suppresses the stop codon (UAG) of the mRNA, the full-length protein obtained upon translation remains attached to the mRNA (read-through ribosome display) that contains the sequence of the tRNA. In this way, the active suppressor tRNAs can be selected (amplified) and their sequences read out. The enriched anticodon (CUA) was complementary to the UAG stop codon and the enriched anticodon-loop was the same as that in the natural tRNASer.

scholarly journals Application of the wheat-germ cell-free translation system to produce high temperature requirement A3 (HtrA3) proteases

BioTechniques ◽  
2012 ◽  
Vol 52 (1) ◽  
Author(s):  
Harmeet Singh ◽  
Shin-ichi Makino ◽  
Yaeta Endo ◽  
Ying Li ◽  
Andrew Stephens ◽  
...  
Keyword(s):  
High Temperature ◽  
Germ Cell ◽  
Wheat Germ ◽  
Translation System ◽  
Temperature Requirement ◽  
Free Translation
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