title: Complementation of argG and hisA mutations of Escherichia coli by DNA cloned from the archaebacterium Methanococcus voltae.
abstract: DNA derived from the methanogenic archaebacterium Methanococcus voltae was digested with PstI restriction endonuclease and cloned into the PstI site of pBR322. The recombinant plasmids generated were used to transform a multiply auxotrophic strain of Escherichia coli with selection for tetracycline resistance. Plasmids complementing the argG(pAW1) or hisA(pAW2) mutations were isolated and characterized. Nick-translated pAW1 and pAW2 hybridized to the predicted M. voltae PstI fragments but not to digested E. coli DNA. A novel 55,000-dalton protein was synthesized in UV-irradiated cells by pAW1, whereas pAW2 synthesized a novel 26,000-dalton protein. Derivatives of pAW1 carrying insertion elements no longer complemented the argG mutation and failed to produce the 55,000-dalton protein. When an AccI fragment was deleted from pAW2, complementation of hisA did not occur and no 26,000-dalton protein was synthesized. The effect of orientation of the cloned DNA within the vector on complementation and polypeptide synthesis was examined.
mesh_terms: D001120:Arginine / Q000096:biosynthesis*; D001426:Bacterial Proteins / Q000096:biosynthesis; D001483:Base Sequence; D003001:Cloning, Molecular; D004269:DNA, Bacterial; D004926:Escherichia coli / Q000235:genetics; D019605:Euryarchaeota / Q000235:genetics*; D005798:Genes, Bacterial; D005816:Genetic Complementation Test; D006639:Histidine / Q000096:biosynthesis*; D009154:Mutation; D010957:Plasmids; D013489:Suppression, Genetic
chemical_list: D001426:Bacterial Proteins; D004269:DNA, Bacterial; D006639:Histidine; D001120:Arginine