Method for determining the sequence, insertion position and marginal sequence of exogenous dna fragments in transgenic organisms

Abstract

The present invention discloses a method for determining the sequence, the insertion position and the boundary sequence of an exogenous DNA fragment in a transgenic organism. The method comprises: sequencing the whole genome of a transgenic organism, comparing a sequence A to the whole genome sequence of the transgenic organism, and determining the sequence, the insertion position and the boundarysequence of the exogenous DNA fragment in the transgenic organism, wherein an acceptor organism is treated by a vector containing the exogenous DNA fragment to obtain the transgenic organism, and thesequence A is the following a1), a2) or a3): a1) the full-length sequence of the vector, a2) the sequence of any one DNA fragment derived from the vector and containing the exogenous DNA fragment, and a3) the sequence of the exogenous DNA fragment or any one fragment thereof. According to the present invention, the method can break through the limitation in species, is not limited by the target gene for constructing the transgenic organism, and can rapidly and precisely detect the insertion position, the boundary sequence and even the copy number of the transgenic gene.

Description

technical field [0001] The invention relates to a method for determining the sequence, insertion position and marginal sequence of a foreign DNA fragment in a transgenic organism in the field of biotechnology. Background technique [0002] In recent years, the safety assessment of genetically modified organisms, especially genetically modified foods, has increasingly become the focus of social attention. Defining the insertion sequence, insertion position and marginal sequence of the transgene in the recipient genome is a key link in evaluating and analyzing the safety of the transgene. Around this problem, people can use the Southern hybridization method to determine the copy number of the transgene, and use methods such as chromosome walking to gradually obtain the insertion position of the transgene in the recipient genome and its marginal sequence. However, these methods are often limited by factors such as transgene copy number, homologous gene similarity, transgene se...

AI Technical Summary

Problems solved by technology

However, these methods are often limited by factors such as transgene copy number, homologous gene similarity, transgene sequence, genome sequence, T-DNA region uncertainty, etc., and the overall efficiency is not high

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