Downstream sequence adjacent to AUG affects translation of chloramphenicol acetyl transferase in eukaryotic cells.

Abstract:

:The CAT gene is widely used as a reporter in eukaryotic systems because of the efficient translation of its mRNA. We report here that a sequence occurring in the CAT mRNA at +15 nucleotides from CAT AUG is essential for translation. This sequence includes a stem-loop structure, which, however, exhibits a calculated stability significantly lower than that required for a hairpin to act as an enhancer of translation in vitro. Replacement of this region with the corresponding sequence from mRNAs that are normally translated in eukaryotic systems drastically reduced translation of CAT in COS cells, although the consensus sequence around the AUG, known to be required for high-level translation initiation, was conserved. These observations may be relevant for the exploitation of the CAT reporter system for analysis of the mechanisms of translation initiation by means of fusion constructs.

journal_name

DNA Cell Biol

journal_title

DNA and cell biology

authors

Ciullo M,Del Pozzo G,Autiero M,Guardiola J

doi

10.1089/104454900314690

keywords:

subject

Has Abstract

pub_date

2000-01-01 00:00:00

pages

39-46

issue

1

eissn

1044-5498

issn

1557-7430

journal_volume

19

pub_type

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