Bacterial Gene Targeting Using Group II Intron Ll.LtrB and the Role of Bacterial Host Factors in Ll.LtrB Splicing and Retrohoming

نویسندگان

  • Jun Yao
  • Alan M. Lambowitz
  • David E. Graham
  • Makkuni Jayaram
  • Richard J. Meyer
  • Ian J. Molineux
  • Junhua Zhao
چکیده

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Retrohoming of a Bacterial Group II Intron Mobility via Complete Reverse Splicing, Independent of Homologous DNA Recombination

The mobile group II intron of Lactococcus lactis, Ll.LtrB, provides the opportunity to analyze the homing pathway in genetically tractable bacterial systems. Here, we show that Ll.LtrB mobility occurs by an RNA-based retrohoming mechanism in both Escherichia coli and L. lactis. Surprisingly, retrohoming occurs efficiently in the absence of RecA function, with a relaxed requirement for flanking ...

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Conserved target for group II intron insertion in relaxase genes of conjugative elements of gram-positive bacteria.

The lactococcal group II intron Ll.ltrB interrupts the ltrB relaxase gene within a region that encodes a conserved functional domain. Nucleotides essential for the homing of Ll.ltrB into an intronless version of ltrB are found exclusively at positions required to encode amino acids broadly conserved in a family of relaxase proteins of gram-positive bacteria. Two of these relaxase genes, pcfG fr...

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Trans-splicing of the Ll.LtrB group II intron in Lactococcus lactis

The Ll.LtrB intron from the Gram-positive bacterium Lactococcus lactis is one of the most studied bacterial group II introns. Ll.LtrB interrupts the relaxase gene of three L. lactis conjugative elements. The relaxase enzyme recognizes the origin of transfer (oriT ) and initiates the intercellular transfer of its conjugative element. The splicing efficiency of Ll.LtrB from the relaxase transcrip...

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Bacterial group II introns and their association with mobile genetic elements.

Group II introns are an abundant class of self-splicing RNAs, found primarily in the organelles of plants and lower eukaryotes and in bacteria. The first bacterial group II intron identified to be functional for splicing in vivo was the Ll.ltrB intron of Lactococcus lactis. It has served as an excellent model for the study of group II intron structure and function. Taking advantage of the tools...

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تاریخ انتشار 2008