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Cloning and characterization of the BglII restriction–modification system reveals a possible evolutionary footprint
- Source :
- Gene. 187:19-27
- Publication Year :
- 1997
- Publisher :
- Elsevier BV, 1997.
-
Abstract
- BglII, a type II restriction–modification (R–M) system from Bacillus globigii, recognizes the sequence 5′-AGATCT-3′. The system has been cloned into E. coli in multiple steps: first the methyltransferase (MTase) gene, bglIIM, was cloned from B. globigii RUB561, a variant containing an inactivated endonuclease (ENase) gene (bglIIR). Next the ENase protein (R.BglII) was purified to homogeneity from RUB562, a strain expressing the complete R–M system. Oligonucleotide probes specific for the 5′ end of the gene were then synthesized and used to locate bglIIR, and the gene was isolated and cloned in a subsequent step. The nucleotide sequence of the system has been determined, and several interesting features have been found. The genes are tandemly arranged, with bglIIR preceding bglIIM. The amino acid sequence of M.BglII is compared to those of other known MTases. A third gene encoding a protein with sequence similarity to known C elements of other R–M systems is found upstream of bglIIR. This is the first instance of a C gene being associated with an R–M system where the R and M genes are collinear. In addition, open reading frames (ORFs) resembling genes involved with DNA mobility are found in close association with BglII. These may shed light on the evolution of the R–M system.
- Subjects :
- DNA, Bacterial
Site-Specific DNA-Methyltransferase (Adenine-Specific)
Molecular Sequence Data
Restriction Mapping
Gene Expression
Bacillus
Biology
Evolution, Molecular
Open Reading Frames
chemistry.chemical_compound
Bacterial Proteins
Escherichia coli
Genetics
DNA Restriction-Modification Enzymes
Amino Acid Sequence
Cloning, Molecular
Deoxyribonucleases, Type II Site-Specific
Peptide sequence
Gene
BglII
DNA Primers
Electrophoresis, Agar Gel
Base Composition
Base Sequence
Oligonucleotide
Nucleic acid sequence
General Medicine
Open reading frame
chemistry
Genes, Bacterial
Restriction modification system
Sequence Alignment
Sequence Analysis
DNA
Subjects
Details
- ISSN :
- 03781119
- Volume :
- 187
- Database :
- OpenAIRE
- Journal :
- Gene
- Accession number :
- edsair.doi.dedup.....c257981ac134bf5ee26a56b25a60260d