The restriction-modification system inStreptomyces flavopersicus

Lyutzkanova, D.; Stoilova−Disheva, Margarita; Peltekova, V.

doi:10.1007/bf02873588

Cited by 3 publications

(4 citation statements)

References 16 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Transformation studies in other bacteria have clearly shown that endogenously modified plasmid DNA can more efficiently transform strains possessing the same R-M system (2,3,15,37,39). The data provided by this study, as well as previous work by other investigators, indicate that a primary limitation to transformation of B. burgdorferi with shuttle vector DNA from E. coli is the barrier presented by the endogenous plasmid-borne R-M systems of B. burgdorferi.…”

Section: Discussionsupporting

confidence: 63%

“…This may indicate that these shuttle vectors do not possess any sites recognized/cleaved by BBH09 or that the enzyme is not active or made in these clones. Transformation studies in other bacteria have shown that methylated plasmid DNA from E. coli can be restricted unless the recipient bacterium contains a similar methyltransferase activity (12,37,38,39,41 (Fig. 3).…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Defining the Plasmid-Borne Restriction-Modification Systems of the Lyme Disease SpirocheteBorrelia burgdorferi

2011

View full text Add to dashboard Cite

The restriction-modification (R-M) systems of many bacteria present a barrier to the stable introduction of foreign DNA. The Lyme disease spirochete Borrelia burgdorferi has two plasmid-borne putative R-M genes, bbe02 and bbq67, whose presence limits transformation by shuttle vector DNA from Escherichia coli. We show that both the bbe02 and bbq67 loci in recipient B. burgdorferi limit transformation with shuttle vector DNA from E. coli, irrespective of its dam, dcm, or hsd methylation status. However, plasmid DNA purified from B. burgdorferi transformed naïve B. burgdorferi much more efficiently than plasmid DNA from E. coli, particularly when the bbe02 and bbq67 genotypes of the B. burgdorferi DNA source matched those of the recipient. We detected adenine methylation of plasmid DNA prepared from B. burgdorferi that carried bbe02 and bbq67. These results indicate that the bbe02 and bbq67 loci of B. burgdorferi encode distinct R-M enzymes that methylate endogenous DNA and cleave foreign DNA lacking the same sequence-specific modification. Our findings have basic implications for horizontal gene transfer among B. burgdorferi strains with distinct plasmid contents. Further characterization and identification of the nucleotide sequences recognized by BBE02 and BBQ67 will facilitate efficient genetic manipulation of this pathogenic spirochete.Borrelia burgdorferi sensu lato is a zoonotic pathogen whose natural infectious cycle alternates between a tick vector and rodent or bird reservoir hosts (1,7,8,14,32,33,36). Transmission of B. burgdorferi to humans occurs through the bite of an infected tick and can lead to Lyme disease, which is a major public health concern in areas of North America and Europe where B. burgdorferi is endemic (8, 53).The genomic structure of the spirochete B. burgdorferi is unique, consisting of a linear chromosome of approximately 900 kb and more than 20 linear (lp) and circular (cp) plasmids, ranging in size from ϳ5 kb to 56 kb, in the type strain B31 (9,10,11,19,42). The plasmids of B. burgdorferi are present at unit copy number relative to the chromosome (22), and some are relatively unstable during in vitro propagation (52, 57). The loss of linear plasmids lp25, lp28-1, and lp36 by strain B31 was found to correlate with the loss of infectivity in mice (20,31,45,56), leading to the identification of genes carried on these plasmids that are dispensable in vitro but required in vivo during an experimental infectious cycle (21,26,35,44,47). The loss of two linear plasmids, lp25 and lp56, was shown to correlate with enhanced shuttle vector transformation, suggesting that specific lp25 and lp56 gene products present a barrier to stable introduction of foreign DNA (34). Further studies linked the transformation phenotype of B. burgdorferi strain B31 with the bbe02 and bbq67 genes on lp25 and lp56, respectively, and the putative restriction-modification (R-M) enzymes that they encode (11,27,29,34). The recent demonstration by Chen and colleagues of enhanced transformation of B. burgdorferi follow...

show abstract

Section: Discussionsupporting

confidence: 63%

Section: Discussionmentioning

confidence: 99%

Defining the Plasmid-Borne Restriction-Modification Systems of the Lyme Disease SpirocheteBorrelia burgdorferi

2011

View full text Add to dashboard Cite

show abstract

“…pogona through genetic and metabolic engineering, which likely requires an efficient transformation system. Bacteria usually possess a restriction–modification (RM) system as part of their defense systems . Previous research indicated that 88% of bacterial genomes contain genes encoding RM system components, with 44% of bacterial genomes encoding four or more RM systems .…”

Section: Introductionmentioning

confidence: 99%

“…Bacteria usually possess a restriction−modification (RM) system as part of their defense systems. 14 Previous research indicated that 88% of bacterial genomes contain genes encoding RM system components, with 44% of bacterial genomes encoding four or more RM systems. 15 On the basis of their subunit composition (i. restriction enzyme and methylase), cofactor requirements, cleavage mode, and other characteristics, RM systems have been divided into four groups (types I−IV).…”

Section: Introductionmentioning

confidence: 99%

Exploring a High-Efficiency Genetic Transformation System for Engineering Saccharopolyspora pogona ASAGF58 To Improve Butenyl-Spinosyn Production

Pang

Guo

et al. 2023

ACS Agric. Sci. Technol.

View full text Add to dashboard Cite

Butenyl-spinosyn is a potent insecticide potentially useful as a broad-spectrum pesticide. Because it is relatively nontoxic to mammals and does not damage the environment, there has been considerable interest in the use of butenyl-spinosyn for increasing agricultural production. However, genetically engineering Saccharopolyspora pogona ASAGF58 to increase its relatively low butenyl-spinosyn content remains challenging because it cannot be transformed efficiently. In this study, genes encoding novel methyltransferases (04455 and 28970) were identified in the Sa. pogona ASAGF58 genome through a bioinformatic-based analysis. Additionally, the transformation efficiency increased by 5.8-and 16.4-fold when foreign DNA was pre-methylated in ET-28970 and ET-04455, respectively, through bypassing of the restriction−modification system. A comparative proteomic analysis of Sa. pogona and Saccharopolyspora spinosa revealed that acetyl-CoA synthetase may be useful for improving butenyl-spinosyn production. The fermentation results indicated that compared with the wild-type butenyl-spinosyn content, overexpressing the acetyl-CoA synthetase gene increased butenyl-spinosyn production by 2-fold. The findings presented herein suggest that the strategy employed in this study may be applicable for the genetic engineering of other nonmodel Saccharopolyspora strains and increase target product yields.

show abstract

Towards a new science of secondary metabolism

2013

View full text Add to dashboard Cite

Secondary metabolites are a reliable and very important source of medicinal compounds. While these molecules have been mined extensively, genome sequencing has suggested that there is a great deal of chemical diversity and bioactivity that remains to be discovered and characterized. A central challenge to the field is that many of the novel or poorly understood molecules are expressed at low levels in the laboratory-such molecules are often described as the 'cryptic' secondary metabolites. In this review, we will discuss evidence that research in this field has provided us with sufficient knowledge and tools to express and purify any secondary metabolite of interest. We will describe 'unselective' strategies that bring about global changes in secondary metabolite output as well as 'selective' strategies where a specific biosynthetic gene cluster of interest is manipulated to enhance the yield of a single product.

show abstract

The restriction-modification system inStreptomyces flavopersicus

Cited by 3 publications

References 16 publications

Defining the Plasmid-Borne Restriction-Modification Systems of the Lyme Disease SpirocheteBorrelia burgdorferi

Defining the Plasmid-Borne Restriction-Modification Systems of the Lyme Disease SpirocheteBorrelia burgdorferi

Exploring a High-Efficiency Genetic Transformation System for Engineering Saccharopolyspora pogona ASAGF58 To Improve Butenyl-Spinosyn Production

Towards a new science of secondary metabolism

Contact Info

Product

Resources

About