Brassica napus BnGLABRA2 promoter and preparation method and application thereof

Abstract

The invention discloses a brassica napus BnGLABRA2 promoter and a preparation method and application thereof. The method comprises the following steps of: A, cloning a promoter sequence by genomic walking, namely extracting genomic DNA (Deoxyribose Nucleic Acid) by using an SDS (Sodium Dodecyl Sulfonate) cracking method, performing genomic walking, and amplifying PCR (Polymerase Chain Reaction) cloned rape twice during each genomic walking; B, performing bioinformatics analysis on the promoter sequence, namely performing homology comparison on a cloned sequence by BLASTN, wherein a promoter sequence and an upstream promoter element are obtained from a promoter core element and an upstream cis acting element by using promoter prediction software; and C, amplifying a full-length promoter and a missing promoter and constructing a pMD18-T-GP vector, namely designing and amplifying a primer of each missing segment by taking the genomic DNA as a template according to the previously obtained promoter sequence for PCR amplification. The application of the brassica napus BnGL2 genomic full-length promoter SEQ ID NO:1 to leaf epidermal hairs, seeds, root parts, entire veins and stele leaf veins is disclosed. The method has the advantages of easiness in operating, stable and reliable result. The brassica napus BnGLABRA2 promoter has biological safety and development and application values.

Description

technical field [0001] The invention relates to the fields of molecular biology and biotechnology, in particular to a BnGLABRA2 promoter of Brassica napus, a preparation method of the BnGLABRA2 promoter of Brassica napus, and the application of the BnGLABRA2 promoter of Brassica napus in genetic engineering of rapeseed. Background technique [0002] Plant gene promoters play a key role in the regulation of gene expression. The regulation of gene expression is the result of the comprehensive action of many factors. Generally, according to the sequence of events, the regulation of genes is divided into regulation at the transcriptional level, regulation at the translational level, and regulation at the protein processing level. The proteins and RNAs encoded by genes and their secondary metabolites are extremely important for maintaining the whole life activities of organisms. Any misregulation of gene expression can have serious consequences for life. Therefore, the researc...

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Problems solved by technology

The application of inducible promoters also has certain limitations. The external condition treatment of recipient plants, such as heat shock, hormone treatment, etc., may cause a series of physiological and biochemical reactions in the organism, which is not conducive to the normal growth of plants.

Moreover, the methyl dehydrocortisol (dex, dexamethasone), estradiol (estradiol) and tetracycline (tetracycline) used as inducers in the chemical regulation system are all harmful to the ecological environment and should not be used in production practice.

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