Target gene trapping method and application thereof

Abstract

The invention relates to a target gene trapping method based on a library and application of the method. The target gene trapping method comprises that genome deoxyribonucleic acid (DNA) fragmentations which are broken into 2-4k are connected with vectors to obtain recombined vectors; the recombined vectors are subjected to electrotransformation and turned into GB05RedTrfA+pSC101Beta competent cells, the competent cells are cultured to obtain a recombined vector bacterium library; double-stranded DNA recombined fragmentations of target genes to be trapped are prepared; and the recombined fragmentations and positive clones in the recombined vector bacterium library are subjected to recombination, and the positive clones are chosen to obtain bacterium clones with target gene fragmentations. The invention further discloses a genome DNA library for target gene trapping. The target gene trapping method and the genome DNA library can be applied to gene study.

Description

technical field [0001] The invention belongs to the field of molecular biology, in particular to a library-based target gene capture method and its application. Background technique [0002] The construction of BAC, PAC, YAC and other cloning vectors provides a powerful tool for gene cloning. In the case of known partial sequence information of the gene, in order to obtain the full-length sequence of the gene, there are mainly two methods in the existing technology. The first feasible method is to use polymerase chain reaction (Polymerase Chain Reaction, PCR) to amplify clones with target fragments from BAC, PAC or YAC libraries; the second method is to use specific sequences to recombine BAC, PAC or YAC library, get positive clones with target fragments. It can be seen that both methods require the construction of BAC, PAC or YAC genomic libraries, but BAC, PAC, YAC library construction and recombination operations are complex, unstable, expensive, long cycle, and cumbers...

AI Technical Summary

Problems solved by technology

However, the improvement of the above-mentioned recombination technology still cannot overcome the shortcomings of the target gene capture technology based on large fragment genome libraries, such as complex operation, high price, and long cycle, which is not convenient for studying the function of small and medium fragment genes.

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