Cotton promoter GbU6-5PS and application

Abstract

The invention provides a cotton promoter GbU6-5PS and application. The positive and negative chains of the cotton promoter GbU6-5PS are as shown in SEQ ID NO.1 and SEQ ID NO.2. The promoter is obtained by conducting the first turn of PCR amplification so as to obtain a segment which covers full-length GbU6-5 promoter sequence; and then, conducting the second turn of PCR amplification, taking the GbU6-5P as a donor plasmid template and AtU6-26SK as an acceptor plasmid template, and truncating and cloning the GbU6-5P promoter by virtue of Transfer PCR method, so that the cotton promoter GbU6-5PS is obtained. The cotton promoter GbU6-5PS disclosed by the invention is not only applicable to cotton, but also quite short in segment which is just 105bp long, conforming to the requirement of a CRISPR / Cas9 genome editing vector, the transcriptional level of sgRNA is significantly improved, and subsequently a genome editing efficiency may be increased.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a cotton promoter GbU6-5PS and its application. Background technique [0002] Genome editing technology is an important tool for studying gene functions. It can precisely modify genes at specific sites on the chromosomes of recipient cells, and can efficiently generate functionally inactive mutants of specific genes, providing high-quality genetic materials for biological functional genome research. . Therefore, this technique has been favored by biologists since its inception. The type II CRISPR / Cas9 genome editing system is another new technology for highly efficient site-directed modification of the genome after zinc finger nucleases (ZFNs) and TALE nucleases (TALENs). sgRNA and Cas9 are two essential elements of this technical system, in which sgRNA plays the role of targeting and binding to the target gene site during the CRISPR / Cas9 genome editing process. In the CRISPR / Cas9...

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Problems solved by technology

[0003] Although the U6 promoter has the characteristics of high-efficiency transcription, it is not necessarily applicable between different species with distant homologous relationships, and there are multiple U6 promoters in the genome of the same species, and the transcription efficiency of these promoters is not the same

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