Cloning and characterization of the BglII restriction–modification system reveals a possible evolutionary footprint
| Autor: | Lucia Greenough, Laurie S. Moran, Joan E. Brooks, Emma Jean Hess, Barton E. Slatko, Daniel F. Heiter, Jack S. Benner, Brian P. Anton |
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| Rok vydání: | 1997 |
| Předmět: |
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 |
| Zdroj: | Gene. 187:19-27 |
| ISSN: | 0378-1119 |
| DOI: | 10.1016/s0378-1119(96)00638-5 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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