We are analyzing https://link.springer.com/article/10.1007/s00203-013-0910-x.

Title:
The tyrosine recombinase MrpA and its target sequence: a mutational analysis of the recombination site mrpS resulting in a new left element/right element (LE/RE) deletion system | Archives of Microbiology
Description:
MrpA is the multimer resolution protein of the Streptomyces coelicolor A3(2) plasmid SCP2*. Previously, MrpA was found to be a site-specific tyrosine recombinase that acts with the 36-bp recombination site mrpS. The present report gives a comprehensive characterization of the composition as well as the position of the spacer and MrpA binding sites within mrpS. Experiments revealed a spacer consisting of 6 remarkably variable nucleotides in the middle of the mrpS-site. A reduction in the spacer to 5 nucleotides abolished recombination. Investigation of the MrpA binding sites showed the importance of its 15 nucleotides on an effective recombination. Among almost randomly exchangeable nucleotides, two nucleotides were identified as essential for MrpA binding. Alteration of either of these nucleotides led to a reduction in MrpA binding down to 2 % or even to no binding. Based on these results, a new left element/right element (LE/RE) deletion system was developed. The constructed heteromeric mrpS-sites are efficiently resolved by MrpA. The resulting double mutated (LE/RE) site can no longer be used as a recombination site by MrpA. The system has been successfully applied for the generation of multiple-targeted deletions in the genome of E. coli.
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Keywords {🔍}

google, scholar, article, cas, pubmed, recombination, dna, recombinase, mrpa, gene, sitespecific, biol, microbiol, site, system, streptomyces, coli, scp, binding, mol, tyrosine, altenbuchner, plasmid, sites, nucleotides, nucleic, acids, res, escherichia, sequence, coelicolor, chem, lox, appl, cloning, acid, privacy, cookies, content, analysis, element, lere, protein, genome, recombinases, cre, mutant, cells, sherratt, proc,

Topics {✒️}

flp-recombinase-mediated cassette-exchange technology multimer resolution protein month download article/chapter replication-recombination-chromosome segregation connection double-digest restriction fragments markerless multiple-gene-deletion system site-specific tyrosine recombinase flp site-specific recombination site-specific recombination synapse recombinase-mediated cassette exchange lac repressor-operator interactions cre-mediated chromosome translocations δ1-pyrroline-5-carboxylic acid site-directed integration system specific dna regions wild-type escherichia coli constructed heteromeric mrps-sites dna cloning system site-specific recombinases loxp-mutated deletion cassettes site-specific recombination site-specific recombinase tyrosine recombinase mrpa cre recombinase protein mrpa binding sites tyrosine recombinase reactions full article pdf streptomyces plasmid scp2 protein domain needed fertility plasmid scp2 privacy choices/manage cookies cold spring harbor article warth resulting double mutated dna recombination multiple-targeted deletions tyrosine recombinases multiple-deletion method cre recombinase complexed cre recombinase resides generating precise deletions analyzing genetic recombination mutant lox sites cryptic lox sites competent escherichia coli escherichia coli k-12 antibiotic-resistance cassettes embryonic stem cells 25 kda carboxyl domain negative selection system

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      description:MrpA is the multimer resolution protein of the Streptomyces coelicolor A3(2) plasmid SCP2*. Previously, MrpA was found to be a site-specific tyrosine recombinase that acts with the 36-bp recombination site mrpS. The present report gives a comprehensive characterization of the composition as well as the position of the spacer and MrpA binding sites within mrpS. Experiments revealed a spacer consisting of 6 remarkably variable nucleotides in the middle of the mrpS-site. A reduction in the spacer to 5 nucleotides abolished recombination. Investigation of the MrpA binding sites showed the importance of its 15 nucleotides on an effective recombination. Among almost randomly exchangeable nucleotides, two nucleotides were identified as essential for MrpA binding. Alteration of either of these nucleotides led to a reduction in MrpA binding down to 2 % or even to no binding. Based on these results, a new left element/right element (LE/RE) deletion system was developed. The constructed heteromeric mrpS-sites are efficiently resolved by MrpA. The resulting double mutated (LE/RE) site can no longer be used as a recombination site by MrpA. The system has been successfully applied for the generation of multiple-targeted deletions in the genome of E. coli.
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         Tyrosine recombinase
         Site-specific recombination
         DNA binding site
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         Microbial Ecology
         Biochemistry
         general
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         Biotechnology
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