Cloning, nucleotide sequence, and expression of the HincII restriction-modification system
| Autor: | Hirokazu Kotani, Teruya Nakamura, Hiroyuki Ito, Atsuko Sadaoka, Nobutsugu Hiraoka |
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| Jazyk: | angličtina |
| Rok vydání: | 1990 |
| Předmět: |
Molecular Sequence Data
Restriction Mapping Molecular cloning Methylation Endonuclease Restriction map Sequence Homology Nucleic Acid Genetics Consensus sequence Amino Acid Sequence Cloning Molecular Deoxyribonucleases Type II Site-Specific Peptide sequence chemistry.chemical_classification biology Base Sequence Adenine Nucleic acid sequence Gene Expression Regulation Bacterial Molecular biology Haemophilus influenzae Amino acid Molecular Weight Blotting Southern chemistry Genes Bacterial biology.protein Restriction modification system Plasmids |
| Popis: | Two genes, coding for the HincII from Haemophilus influenzae Rc restriction-modification system, were cloned and expressed in Escherichia coli RR1. Their DNA sequences were determined. The HincII methylase (M.HincII) gene was 1,506 base pairs (bp) long, corresponding to a protein of 502 amino acid residues (Mr = 55,330). The HincII endonuclease (R.HincII) gene was 774 bp long, corresponding to a protein of 258 amino acid residues (Mr = 28,490). The amino acid residues predicted from the R.HincII and the N-terminal amino acid sequence of the enzyme found by analysis were identical. These methylase and endonuclease genes overlapped by 1 bp on the H. influenzae Rc chromosomal DNA. The clone, named E. coli RR1-Hinc, overproduced R.HincII. The R.HincII activity of this clone was 1,000-fold that from H. influenzae Rc. The amino acid sequence of M.HincII was compared with the sequences of four other adenine-specific type II methylases. Important homology was found between tne M.HincII and these other methylases. |
| Databáze: | OpenAIRE |
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