Molecular cloning and sequencing of mcrA locus and identification of McrA protein in Escherichia coli.
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Date
1992-09
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Abstract
The Mcr systems (previously known as Rgl systems) of Escherichia coli
recognize and cleave specific sequences carrying methylated or hydroxymethylated
cytosines. We have cloned the mcrA gene and determined its nucleotide sequence. An 831
base pair sequence encodes the McrA protein. Analysis of the sequence data reveals that
there arc additional ORFs internal to the above. A phage T7 expression system was used
to determine the protein products encoded by the cloned mcrA gene. The results clearly
show that a 31 kDa polypeptide is responsible for McrA activity. This is in agreement with
the molecular weight deduced from sequence data. McrA protein was found to be localized
in the outer membrane of Escherichia coli. To our knowledge this is the first restriction
enzyme localized in the outer membrane of Escherichia coli.
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Keywords
mcrA, el4, mcrA sequence, T7 expression system
Citation
Ramalingam R, Prasad Ranjan, Shivapriya R, Dharmalingam K. Molecular cloning and sequencing of mcrA locus and identification of McrA protein in Escherichia coli. Journal of Biosciences. 1992 Sep; 17(3): 217-232.