Process for cloning DNA sequences or for the production of gene libraries, cloning vectors, and micro-organism strains containing them

摘要

The process entails a) removing from the cohesive 5' ends of a cloning vector, which has been isolated as linear DNA after treatment with restriction endonuclease, the phosphate groups with phosphatase, adding DNA fragments which contain the gene to be cloned, have been generated by treatment with restriction endonuclease and have phosphate groups on their ends to the fragments which are incapable of joining together, and ligating the mixture with ligase, or b) cutting a recombinant cloning vector of the phage, cosmid, phasmid or transphasmid type, containing cohesive sequences, with terminase, and carrying out in vitro assembly (packaging) of the monomer DNA obtained by one of the above alternatives, and then replicating the phages.