Mutation carrier and application

Abstract

A mutation carrier able to clone DNA to it for quickly completing mutation and its applicatino are disclosed. Its advantages are simple and reliable application, and no pollution to original template.

Description

Technical field [0001] The invention relates to a DNA mutation tool in genetic engineering technology, in particular to a mutation vector capable of cloning DNA onto it to complete mutation concisely and quickly and its application. Background technique [0002] There are many ways to mutate genes. End overlap PCR is a commonly used one; Ho et al. used this method as early as 1989 to realize the cutting, joining and mutation of genes (Ho SN, Hunt HD, Horton RM. et al Site directed mutagenesis by overlap extension using the polymerase chain reaction. Gene 1989; 77: 51-59.). In 1997, Warrens et al. improved this mutation method (Warrens AN, Jones MD, Lechler RI. Splitting by overlap extension by PCR using asymmetric amplification: an improved technique for the generation of hybrid proteins of immunological interest. Gene. 1997 20; 186 (1): 29-35.). Until recently, this was still one of the common methods of gene mutation; but it has the disadvantage of complicated operation. In act...

AI Technical Summary

Problems solved by technology

Unmutated products account for the vast majority of the final PCR products, making subsequent screening cumbersome

Urban, Xu, Wang, Li etc. have improved this method, but their preliminary work is also more complicated, and some will use the expensive DpnI enzyme to remove pollution (Urban A, Neukirchen S, Jaeger KE.A rapid and efficient method for site-directed mutagenesis using one-step overlap extension PCR. Nuclec Acid Res. 1997 25(11): 2227-2228 Xu W, Zhang Y, Yeh LY et al. One-step, highly efficient site-directed mutagenesis by toxic protein selection .Biotechniques.2002 32(6):1266-1268, 1270. Wang W, Malcolm BA. Two-stage polymerase chain reaction protocol allowing introduction of multiple mutations, deletions, and insertions, using QuikChange site-directed mutagenesis.Methods2 Mol02 Biol. 182: 37-43. Li F, Mullins JI. Site-directed mutagenesis facilitated by DpnI selection on hemimethylated DNA. Methods Mol Biol. 2002; 182: 19-27.)

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