Simplified in vitro system for study of eukaryotic mRNA translation by measuring di- and tripeptide formation.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 80 ·No. 11 ·1983-06-00 ·Pages 3223-6

Cenatiempo Y, Twardowski T, Redfield B, Reid BR, Dauerman H, Weissbach H, Brot N

Abstract

An in vitro system for measurement of rabbit globin mRNA translation has been developed based on the formation of the NH2-terminal dipeptide, fMet-Val. The basic components include a partially purified initiation factor preparation from rabbit reticulocytes supplemented with eukaryotic initiation factor 4A, purified and formylated yeast Met-tRNAi, and rabbit liver or Escherichia coli Val-tRNA1Val. Picomole quantities of fMet-Val are synthesized, dependent on mRNA, and the dipeptide is readily assayed by a simple extraction procedure. In the presence of Leu-tRNA or His-tRNA, the tripeptides fMet-Val-Leu and fMet-Val-His are synthesized, corresponding to the NH2-terminal sequence of alpha- and beta-globin, respectively. Therefore, tripeptide synthesis provides a simple means to distinguish between the expression of the alpha- and beta-globin mRNA species.

MeSH Terms

Animals Artemia Cell-Free System Embryo, Nonmammalian Female Globins/genetics Kinetics Oligopeptides/genetics Protein Biosynthesis RNA, Messenger/genetics RNA, Transfer/genetics Rabbits Ribosomes/metabolism

Chemicals

Oligopeptides RNA, Messenger Globins RNA, Transfer

Authors & Affiliations

7 authors, click to expand affiliations / ORCID

Cenatiempo Y

Twardowski T

Redfield B

Reid B R

Dauerman H

Weissbach H

Brot N

References (21)

21 references, click to expand

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Article Info

Journal

Proceedings of the National Academy of Sciences of the United States of America

Abbr.

Proc Natl Acad Sci U S A

ISSN

0027-8424

DOI

10.1073/pnas.80.11.3223

Published

1983-06-00

Pages

3223-6

Language

English

Region

United States

NLM ID

7505876

PMCID

PMC394012

Subset

External Links

PubMed source DOI: 10.1073/pnas.80.11.3223 PMC Full Text View journal details

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Simplified in vitro system for study of eukaryotic mRNA translation by measuring di- and tripeptide formation.

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