METHOD FOR PREPARING RECOMBINANT DNA MOLECULE FROM ONE OR MORE SINGLE-STRANDED OR DOUBLE-STRANDED MATRIX POLYNUCLEOTIDES (VARIANTS) Russian patent published in 2003 - IPC

FIELD: biotechnology, genetic engineering. SUBSTANCE: method of mutagenesis and recombination of polynucleotide sequences in vitro is based on elongation of primer oligonucleotides catalyzed by polymerase. Method involves priming matrix polynucleotide (polynucleotides) with primers with random sequences or well- defined sequences to provide pool of short DNA fragments with low level of point mutations. DNA fragments are subjected for denaturation followed by annealing and the following DNA polymerization catalyzed by enzyme. Procedure is repeated sufficiently times to obtain full-length genes that comprise mutants of the parent matrix polynucleotides. Then genes amplified using polymerase chain reaction and cloned into vector for expression of encoded proteins. Invention provides preparing gene products with improved properties. Invention can be used for realization of mutagenesis and recombination of polynucleotides sequences in vitro. EFFECT: improved preparing method. 62 cl, 17 dwg, 5 tbl