Convenient and rapid kit of plant gene editing tool p-XCHBCC3

Abstract

The invention relates to a convenient and rapid kit development of a general plant gene editing tool p-XCHBCC3. The preparation method comprises the following steps that firstly, on the basis of an original general vector pKI1.1R, a modified herbicide-resistant Bar gene without a PaqCI enzyme digestion recognition site is added to construct an intermediate vector p-XCHB2; on the basis of the p-XCHB2, two selection markers, namely a ccdB escherichia coli suicide gene and a CmR chloramphenicol resistance gene, are added to an insertion position sequence seat of an empty vector target spot, and a final target empty vector p-XCHBCC3 is prepared. In addition, specific laboratory usage and commercial usage of the editing tool are further described. The kit has the advantages that as a general plant gene editing tool, various defects of some vectors circulating in the current market can be overcome, various advantages of the vectors can be integrated, the operation is simple, the process is rapid, the scientific research workload can be greatly reduced, great benefits are brought to basic research and application research, and the kit has good popularization prospects and value.

Description

technical field [0001] The invention relates to a convenient and rapid plant gene editing tool p-XCHBCC3 kit. Background technique [0002] CRISPR-Cas9 is the most advanced gene editing system at this stage. Using this system, the modification of genetic information precisely manipulated by humans has been realized in many species, which has greatly promoted the development of basic science and the advancement of fields such as medicine and breeding. As a model organism, Arabidopsis is the most perfect material for genetic research in fields such as botany. Several CRISPR-Cas9 editing vectors for Arabidopsis are currently available, and while they help us in our work, each has its own disadvantages: for example, they either need to be used in conjunction with other vectors, or the limitations required Sexual endonucleases are too expensive, or the enzyme cutting efficiency is not high, resulting in unsatisfactory ligation and clone screening difficulties, or the expression...

AI Technical Summary

Problems solved by technology

Several CRISPR-Cas9 editing vectors for Arabidopsis are currently available, and while they help us in our work, each has its own disadvantages: for example, they either need to be used in conjunction with other vectors, or the limitations required Sexual endonucleases are too expensive, or the enzyme cutting efficiency is not high, resulting in unsatisfactory ligation and clone screening difficulties, or the expression effect is not ideal, or only hygromycin or kanamycin can be used as a screening agent It leads to cumbersome screening of seedlings, or it is not easy to screen from the T2 progeny in a short period of time to the offspring of plants that are both homozygous edited individuals and do not contain large exogenous DNA insertions (T-DNA insertions)

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