HaeIII和HaeII甲基转移酶基因的克隆与分析

Cloning and analysis of the HaeIII and HaeII methyltransferase genes.

作者信息

Slatko B E, Croft R, Moran L S, Wilson G G

机构信息

New England Biolabs, Inc., Beverly, MA 01915.

出版信息

Gene. 1988 Dec 25;74(1):45-50. doi: 10.1016/0378-1119(88)90248-x.

DOI:10.1016/0378-1119(88)90248-x

PMID:3248732

Abstract

The HaeIII methyltransferase (MTase) gene from Haemophilus aegyptius (recognition sequence: 5'-GGCC-3') was cloned into Escherichia coli in the plasmid vector pBR322. The gene was isolated on a single EcoRI fragment and on a single HindIII fragment. Clones carrying additional adjacent fragments were found to code also for the HaeII restriction endonuclease and HaeII modification MTase (recognition sequence: 5'-PuGCGCPy-3'). The sequence of the HaeIII modification gene was determined. The inferred amino acid sequence of the protein was found to share extensive similarity with other sequenced m5C-MTases. The central 'non-conserved' region of the M.HaeIII MTase, thought to form the nucleotide sequence-specificity domain, is almost identical to that of the M.BsuRI, M.BspRI and M.NgoPII MTases, which also recognize the sequence 5'-GGCC-3'.

摘要

来自埃及嗜血杆菌的HaeIII甲基转移酶（MTase）基因（识别序列：5'-GGCC-3'）被克隆到质粒载体pBR322中的大肠杆菌中。该基因在单个EcoRI片段和单个HindIII片段上分离得到。发现携带额外相邻片段的克隆也编码HaeII限制内切酶和HaeII修饰MTase（识别序列：5'-PuGCGCPy-3'）。测定了HaeIII修饰基因的序列。发现该蛋白质的推断氨基酸序列与其他已测序的m5C-MTases具有广泛的相似性。M.HaeIII MTase的中心“非保守”区域，被认为形成核苷酸序列特异性结构域，与M.BsuRI、M.BspRI和M.NgoPII MTases的该区域几乎相同，它们也识别序列5'-GGCC-3'。