Cloning and expression of the Hpal restriction-modification genes

descriptionPublicationkeyboard_double_arrow_right Article 01 Jan 1992 English Publisher:Oxford University Press (OUP)Journal:Nucleic Acids Research, volume 20, pages 705-709 (issn: 0305-1048, eissn: 1362-4962,

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Authors: H, Ito; H, Shimato; A, Sadaoka; H, Kotani; F, Kimizuka; I, Kato;

doi: 10.1093/nar/20.4.705

pmid: 1542567

pmc: PMC312008

Cloning and expression of the Hpal restriction-modification genes

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Abstract

The genes from Haemophilus parainfluenzae encoding the HpaI restriction-modification system were cloned and expressed in Escherichia coli. From the DNA sequence, we predicted the HpaI endonuclease (R.HpaI) to have 254 amino acid residues (Mr 29,630) and the HpaI methyltransferase (M.HpaI) to have 314 amino acid residues (37,390). The R.HpaI and M.HpaI genes overlapped by 16 base pairs on the chromosomal DNA. The genes had the same orientation. The clone, named E. coli HB101-HPA2, overproduced R.HpaI. R.HpaI activity from the clone was 100-fold that from H. parainfluenzae. The amino acid sequence of M.HpaI was compared with those of other type II methyltransferases.

Keywords

Site-Specific DNA-Methyltransferase (Adenine-Specific), Base Sequence, Molecular Sequence Data, Restriction Mapping, Haemophilus, Gene Expression, Blotting, Southern, Amino Acid Sequence, Cloning, Molecular, Deoxyribonucleases, Type II Site-Specific

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