Brassica napus ALS (Acetolactate Synthase) gene promoter and application

A Brassica napus and promoter technology, applied in application, genetic engineering, recombinant DNA technology, etc., to achieve the effect of reducing biosafety risks

Inactive Publication Date: 2017-12-12
JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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  • Brassica napus ALS (Acetolactate Synthase) gene promoter and application
  • Brassica napus ALS (Acetolactate Synthase) gene promoter and application
  • Brassica napus ALS (Acetolactate Synthase) gene promoter and application

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[0028] 1. Promoter sequence cloning and promoter element analysis

[0029] The present invention is based on the ALS of rape M342 3 Gene sequence, use NCBI website (https: / / www.ncbi.nlm.nih.gov / ) to retrieve the corresponding rapeseed genome sequence and extract ALS 3 A sequence of approximately -2000 bases upstream of the 5'end of the gene coding region. Use a professional website (http: / / molbiol-tools.ca / Promoters.htm) for promoter prediction and analysis. According to the analysis results, select rich promoter elements Design specific amplification primers for the sequence about -1000 bases upstream. Upstream primers are mainly ALS 3 The upstream sequence of the gene is used as a template, and the downstream primer is M342 ALS 3 The 5'end non-coding region sequence of the gene is used as a template to amplify ALS 3 The upstream of the gene has a DNA sequence of about 1048bp.

[0030] 1.1 Promoter sequence amplification and cloning: Use herbicide-resistant rapeseed M342 (known an...

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Abstract

The invention relates to a brassica napus ALS (Acetolactate Synthase) gene promoter and application and belongs to the technical field of biology. The brassica napus ALS gene promoter disclosed by the invention is characterized in that a brassica napus ALS gene promoter sequence is colonized; a promoter element contained in the sequence is predicted by bioinformatics software; the activity of the gene promoter is subjected to qualitative detection and analysis by constructing a plant expression vector; and the results show that the colonized sequence can drive a GUS reporter gene to be expressed in all organs of tobacco seedlings, has high expression activity and characteristics of a constitutive promoter and can be applied to plant genetic engineering research as the strong constitutive expression promoter. The promoter can replace a cauliflower mosaic virus (CaMV) 35S promoter and an agrobaeterittm tumefaciens nopaline gene nos promoter which are normally used in the current plant genetic engineering research to construct a plant transgenic expression vector; and the plant transgenic expression vector is applied to genetic engineering of dicotyledons, so that the biological safety risk of transgenic plants is reduced.

Description

technical field [0001] The invention relates to a brassica napus ALS gene promoter and its application, and belongs to the field of biotechnology. Background technique [0002] Rapeseed is one of the most important oil crops in the world, and it is also an important oil crop in my country. my country's rapeseed planting area and total output both rank first in the world. In the 1960s and 1970s, the annual planting area of ​​rapeseed in my country was about 30 million mu, with a yield of 30-40 kg per mu; in the 1990s, it had grown to 100 million-110 million mu, with a yield of 90-100 kg per mu, and the total rapeseed output reached 1100- More than 12 million tons. Rapeseed is the fifth largest crop in my country after rice, wheat, corn, and soybean (it will surpass soybean soon, and rapeseed is the second largest crop after rice in provinces such as Hubei in the Yangtze River Basin), and it is also the only winter oil crop. my country's rapeseed area and total output accoun...

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Application Information

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IPC IPC(8): C12N15/113C12N15/82A01H5/00
CPCC12N9/1022C12N15/8222C12Y202/01006
Inventor 陈松浦惠明彭琦张洁夫陈锋王晓东张维胡茂龙龙卫华高建芹付三雄周晓婴熊冬琴
Owner JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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