Tobacco germ cell specific high-expression gene and application thereof

Abstract

The invention relates to a tobacco germ cell specific high-expression gene and application thereof. A nucleotide sequence is shown as SEQ ID NO.1. The coding sequence and the promoter sequence of the gene of cultivated tobacco are obtained through genome informatics analysis and identification. Candidate promoter sequence element analysis and gene expression profile analysis prove that the gene is a germ cell high-abundance expression gene. A promoter sequence of the gene is obtained through cloning, a gene editing vector of germ cell specific high-abundance expression Cas 9 protein is constructed by taking the promoter sequence as a starting element, the vector is transferred into agrobacterium, a transgenic tobacco plant is obtained through a leaf disc conversion method, and gene editing target site sequencing analysis finds that the transgenic tobacco plant PDS gene mutation efficiency is high; the research result has important guiding significance and application value for constructing a high-efficiency plant gene inactivation vector and obtaining a mutant plant.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a specific high-expression gene of tobacco germ cells and its application. Background technique [0002] The principle of the CRISPR / Cas9 system is to recognize the target site through the sgRNA (single-guideRNA, sgRNA) formed by crRNA and tracrRNA, and guide Cas9 to perform directional cutting of the target gene. Cas9 is composed of two domains, HNH and RuvC, The two domains are respectively responsible for cutting the two strands of DNA, forming a DNA double-strand break (DSB). The DSB is mainly repaired by two methods. One is non-homologous end joining (Nonhomologous End Joining, NHEJ), this repair method will cause the insertion and deletion of bases, resulting in the loss of gene function. When there is a donor DNA template, the precise repair mode of homologous recombination (HR) will occur, but its efficiency is low. The CRISPR / Cas9 gene editing system is simple in design, ...

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Problems solved by technology

[0005] The purpose of the present invention is to provide a tobacco germ cell-specific expression gene and its application, to solve the existing CRISPR / Cas9 gene editing vector using conventional 35S as a promoter When editing, the proportion of chimerism in offspring plants is high and the editing efficiency is low, which consumes a lot of manpower, material resources and financial resources.

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