New gene mutation recombination method and application thereof

• Main Authors: YANG QINGJUN, LI NIAN, ZHANG YIZHENG, WAN MINYUAN, FENG HONG
• Format: Patent
• Language: Chinese; English
• Published: 16.09.2009
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMBINATORIAL CHEMISTRY; ENZYMOLOGY; FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIREDCHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERSFROM A RACEMIC MIXTURE; LIBRARIES, e.g. CHEMICAL LIBRARIES, IN SILICOLIBRARIES; METALLURGY; MICROBIOLOGY; MUTATION OR GENETIC ENGINEERING; SPIRITS; VINEGAR; WINE

The invention belongs to the field of gene engineering and relates to a gene mutation recombination method and application thereof in construction of protein (proteinase) gene mutation library. In a polymerase chain reaction (PCR) system for amplifying a target sequence, nucleotide analogue 2-deoxyinosine-5-triphosphoric acid (dITP) is randomly doped in a target gene order by a PCR reaction so as to generate multi-point random mutation, the endonuclease V (endo V) cutting the dITP is distinguished by specificity so as to segment the amplified target gene order, and then the obtained the mutation target gene segments are extended and recombined by the PCT without a primer, and finally the mutation gene library of the recombined mutation gene is obtained. The method of the invention overcomes the shortage that the traditional DNA shuffling method is not easily controlled and wastes time and labour in the process of randomly cutting the target sequence by endonuclease (DNase I). The method can be widely applied