Elsevier

Gene

Volume 74, Issue 1, 25 December 1988, Pages 15-21
Gene

Overproduction and purification of the M·HhaII methyltransferase from Haemophilus haemolyticus

https://doi.org/10.1016/0378-1119(88)90240-5Get rights and content

Abstract

The HhaII methyltransferase gene from Haemophilus haemolyticus was subcloned in an expression vector under control of the hybrid trp-lac promoter. Induction with isopropyl-β-d-thiogalactopyranoside results in overproduction of the methyltransferase to about 3% of total cellular protein. The methyltransferase was purified to near electrophoretic homogeneity by phosphocellulose, DEAE, and gel chromatography. Its monomer Mr by sodium dodecyl sulfate-polyacrylamide gel electrophoresis is 25 kDa, in good agreement with that predicted from the nucleotide sequence. Crystals of the methyltransferase were obtained in the presence of a two-fold molar excess of the duplex oligodeoxynucleotide substrate 5′d-GGACTCCCCTGAGG.

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