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Category: biotechnology & health
BSA 07-32: Methyl CpG Detection using McrA
BNL Reference Number: BSA 07-32
Patent Status: U.S. Patent Number 8,361,746 was issued on January 29, 2013
Summary
Differential binding of Escherichia coli McrA protein to DNA sequences that contain the dinucleotide m5CpG.
An expression vector for the production of full-length active McrA protein for use in isolating and selecting sequences containing 5'-CMeCGG-3' has been invented. Cells containing the expression vector and methods for expressing and isolating highly purified McrA protein are provided.
Description
The complete gene for the McrA protein has been cloned in a bacterial expression vector. The expression of the protein in bacterial cells is enhanced by the presence of plasmids containing suitable tRNA genes. Purification of the expressed protein is facilitated by expression, in-frame, of various affinity tags.
Benefits
The determination of the methylation state of specific gene sequences is facilitated by an array of methyl-binding proteins each having specific recognition capacities. The use of the binding proteins facilitates direct analysis of the methylation state and is less cumbersome than methods involving bisulfite sequencing or methyl-sensitive restriction analysis.
Applications and Industries
The development of diagnostic and prognostic systems enabling the determination of the methylation state of various gene sequences depends on having a complete set of tools. The McrA protein has a sequence recognition capability that ideally complements methyl-binding proteins that are currently available.
Have Questions?
For more information about this technology, contact Christine Brakel, (631) 344-7134.
Tags: CpG methylation, DNA assay
