Sain B, Murray N E
Mol Gen Genet. 1980;180(1):35-46. doi: 10.1007/BF00267350.
The hsd genes of E. coli K 12 have been cloned in phage lambda by a combination of in vitro and in vivo techniques. Three genes, whose products are required for K-specific restriction and modification, have been identified by complementation tests as hsdR, M and S. The order of these closely linked genes was established as R, M, S by analysis of the DNA of genetically characterised deletion derivatives of lambda hsd phages. The three genes are transcribed in the same direction but not necessarily as a single operon. Genetic evidence identifies two promoters, one from which transcription of hsdM and S is initiated and a second for the hsdR gene. The hsdR gene codes for a polypeptide of molecular weight approximately 130 000; hsdM for one of 62--65 000 and the hsdS gene was associated with two polypeptides of approximately 50 000. Circumstantial evidence suggest that one of these two polypeptides may be a degradation, or processed, derivative of the other. The hsdS polypeptide of E. coli B has a slightly higher mobility in an SDS-polyacrylamide gel than does that of E. coli K 12. A probe comprising most of the hsdR gene and all of the hsdM and S genes of E. coli K 12 shares extensive homology with the DNA of E. coli B but none with that of E. coli C.
通过体外和体内技术相结合的方法,已将大肠杆菌K12的hsd基因克隆到噬菌体λ中。通过互补试验鉴定出三个基因,其产物是K特异性限制和修饰所必需的,分别为hsdR、M和S。通过分析λhsd噬菌体的遗传特征缺失衍生物的DNA,确定了这些紧密连锁基因的顺序为R、M、S。这三个基因按相同方向转录,但不一定作为单个操纵子转录。遗传学证据确定了两个启动子,一个启动hsdM和S的转录,另一个启动hsdR基因的转录。hsdR基因编码一种分子量约为130000的多肽;hsdM编码一种分子量为62 - 65000的多肽,hsdS基因与两种分子量约为50000的多肽相关。间接证据表明,这两种多肽中的一种可能是另一种的降解产物或加工衍生物。大肠杆菌B的hsdS多肽在SDS - 聚丙烯酰胺凝胶中的迁移率略高于大肠杆菌K12的hsdS多肽。一个包含大肠杆菌K12大部分hsdR基因以及所有hsdM和S基因的探针与大肠杆菌B的DNA具有广泛的同源性,但与大肠杆菌C的DNA没有同源性。