Sequencing and expression of the rne gene of escherichia coli

Anil K. Chauhan, Andras Miczak, Laimute Taraseviciene, David Apirion

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Abstract

RNase E is a major endonucleolytic RNA processing enzyme in Escherichia coli. We have sequenced a 3.2 kb EcoRI-SamHI fragment encoding the me gene, and identified its reading frame. Upstream from the gene, there are appropriate consensus sequences for a putative promoter and a ribosome binding site. We have translated this gene using a T7 RNA polymerase/promoter system. We determined 25 amino acids from the N-terminal of the translated product and they are in full agreement with the DNA sequence. The translated product of the me gene migrates in SDS containing polyacrylamide gels as a 110,000 Da polypeptlde, but the open reading frame found In the sequenced DNA Indicates a much smaller protein. The entity that migrates as a 110,000 Da contains RNA, which could account, at least partially, for the migration of the me gene product In SDS containing polyacrylamide gels.

Original languageEnglish
Pages (from-to)125-129
Number of pages5
JournalNucleic acids research
Volume19
Issue number1
DOIs
Publication statusPublished - Jan 11 1991

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ASJC Scopus subject areas

  • Genetics

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