Method for cleaning DNA transformation plant without choosing marker gene

Abstract

The invention discloses a method for cleaning a DNA transformation plant without choosing a marker gene. The method comprises the following steps of: 1), preparing a target gene expression frame: adopting an appropriate restriction enzyme to cut a constructed target gene expression frame DNA fragment from a plasmid vector according to the sequence character of the plasmid vector on which the target gene expression frame to be transferred is, and obtaining the target gene expression frame DNA by electrophoresing, separating, tapping and reclaiming a gel; 2), preparing a DNA particle vector suspension; 3), bombarding plant callus to transform the plant callus; 4), regenerating the plant; and 5), choosing transgenic plants. When the method is adopted to perform plant transgenosis, the influences of the selected marker gene and superfluous DNA sequences such as the plasmid vector frame DNA sequence on the transgenic plants and genetically modified foods can be overcome fundamentally, so that the transgenic plants without other superfluous DNAs except for the target gene expression frame are obtained.

Description

technical field The invention relates to a method for transforming plants using a gene gun to mediate an exogenous gene expression frame (promoter-gene coding region-terminator). This method does not need to use any selection marker gene, and the obtained transgenic plants have no Contains no excess junk DNA, is a clean DNA transformation technology (Clean DNA transformation). Belongs to the field of biology, especially related to genetic engineering technology. Background technique Since the birth of the first transgenic plant in 1983, plant gene introduction technology has been perfected day by day, and a plant gene transformation system based on two major transformation technologies of gene gun and Agrobacterium has been formed. Dozens of transgenic plants including soybean, corn, rapeseed, cotton, potato, rice and other major crops have entered field trials and even commercialized planting in many countries. At the same time, the potential ecological and environmental ...

AI Technical Summary

Problems solved by technology

Patent publication date: on July 3rd, 2002, patent publication number CN 1356389A), but this method realizes gene transformation by the homologous exchange recombination between the conserved sequence of the plant nucleus DNA that is set at the transformation fragment two ends and plant nucleus DNA, this The defect of this method is: in addition to the target gene, additional plant DNA conservative sequences must be introduced at the two segments of the transformed DNA fragment, and these additional plant DNA conservative sequences are not only a kind of extra redundant junk DNA for the transformation recipient plant, but also Since the conservative sequence of the plant DNA that promotes transformation is set based on the conserved sequences on both sides of the structural gene encoding the protein in higher plants, when the exogenous target gene is integrated into the plant genome through homologous recombination, it will inevitably destroy the recipient plant. The original structural genes lead to genetic mutations in transgenic recipient plants, which lead to new safety risks

Images