Expression of the mcrA gene of escherichia coli is regulated posttranscriptionally, possibly by sequestration of the Shine-Dalgarno region

Shivapriya, R. ; Prasad, Ranjan ; Narayanan, Iyer Lakshmi ; Krishnaswamy, S. ; Dharmalingam, K. (1995) Expression of the mcrA gene of escherichia coli is regulated posttranscriptionally, possibly by sequestration of the Shine-Dalgarno region Gene, 157 (1-2). pp. 201-207. ISSN 0378-1119

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Official URL: http://www.sciencedirect.com/science/article/pii/0...

Related URL: http://dx.doi.org/10.1016/0378-1119(94)00746-F

Abstract

The polypeptides encoded by the mcrA gene were analysed using a T7 expression system. Cloned fragments of 1.6 and 1.0 kb displayed an McrA+/Rg1A+ phenotype and directed synthesis of a 31-kDa polypeptide. A derivative of these clones altered at an internal HindIII site displayed an McrA+/Rg1A+ phenotype and directed production of a 23-kDa polypeptide. Smaller inserts displayed McrA-/Rg1A- phenotypes, though a 0.7-kb insert did direct production of a 24-kDa polypeptide. A construct carrying the 1.0-kb mcrA insert yielded a single 1.3-kb transcript. The mcrA transcript was found to start from C, G, T and G, namely the fourth, fifth, sixth and seventh nucleotides (nt), respectively, downstream from the last nt of the putative - 10 region. Two mcrA transcriptional/transational fusions were made in the pT7-7 expression vector and the protein encoded by these constructs were analysed. Regulation of mcrA expression was studied by quantitative Northern analysis of RNA from various mcrA clones. Together with a computer analysis of the translation initiation region in these mRNAs, the results suggest that the expression of mcrA may be regulated at the translational level.

Item Type:Article
Source:Copyright of this article belongs to Elsevier Science.
Keywords:S1 Analysis; Primer Extension; Overexpression; Regulation
ID Code:70419
Deposited On:17 Nov 2011 03:45
Last Modified:17 Nov 2011 03:45

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