Open AccessCloning and expression of the Hpal restriction-modification genes
Author(s) -
Hiroyuki Ito,
Harumi Shimato,
Atsuko Sadaok,
Hirokazu Kotani,
Fusao Kimizuka,
Ikuo Kato
Publication year - 1992
Publication title -
nucleic acids research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 9.008
H-Index - 537
eISSN - 1362-4954
pISSN - 0305-1048
DOI - 10.1093/nar/20.4.705
Subject(s) - biology , restriction enzyme , gene , clone (java method) , cloning (programming) , microbiology and biotechnology , molecular cloning , peptide sequence , nucleic acid sequence , genetics , escherichia coli , dna , restriction map , endonuclease , methyltransferase , amino acid , methylation , computer science , programming language
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.