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PMID: 2374714 Published · ppublish English Journal Article

Cloning, nucleotide sequence, and expression of the HincII restriction-modification system.

Nucleic acids research ·Vol. 18 ·No. 13 ·1990-07-11 ·Pages 3903-11

Ito H, Sadaoka A, Kotani H, Hiraoka N, Nakamura T

Abstract

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.

MeSH Terms
Adenine/metabolism Amino Acid Sequence Base Sequence Blotting, Southern Cloning, Molecular Deoxyribonucleases, Type II Site-Specific/biosynthesis,genetics,metabolism Gene Expression Regulation, Bacterial Genes, Bacterial Haemophilus influenzae/enzymology,genetics Methylation Molecular Sequence Data Molecular Weight Plasmids Restriction Mapping Sequence Homology, Nucleic Acid
Chemicals
Deoxyribonucleases, Type II Site-Specific GTYRAC-specific type II deoxyribonucleases Adenine
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Ito H
Bioproducts Development Center, Takara Shuzo Co., Ltd., Shiga, Japan.
Sadaoka A
Kotani H
Hiraoka N
Nakamura T
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/18.13.3903
Published
1990-07-11
Pages
3903-11
Language
English
Region
England
NLM ID
0411011
PMCID
PMC331092
Subset
IM
Databases
GENBANK
X52124
External Links
PubMed source DOI: 10.1093/nar/18.13.3903 PMC Full Text View journal details
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