Application of soybean gene promoters pRPS28 and pRPS28-I in soybeans, arabidopsis thaliana and tobaccos

A prps28-i, promoter technology, applied in application, genetic engineering, plant gene improvement and other directions, can solve the problems of gene silencing, imperfect screening conditions for constitutive promoters, and no involvement of soybean root nodule development and expression.

Active Publication Date: 2021-10-01
HENAN UNIVERSITY
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  • Application Information

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Problems solved by technology

In addition, in terms of screening methods, the screening conditions of the reported constitutive promoters are mostly incomplete. Most of the promoters are obtained from the tissue expression in a single period of soybean growth and development, while the constitutive expression in soybean root nodule development The promoter of the
In terms of screening technology, most of the reported promoters are detected by the traditional qRT-PCR method, and there are not many reports on the detection of expression levels by high-throughput RNA sequencing methods
Literature (Ning Zhang, Leah

Method used

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  • Application of soybean gene promoters pRPS28 and pRPS28-I in soybeans, arabidopsis thaliana and tobaccos
  • Application of soybean gene promoters pRPS28 and pRPS28-I in soybeans, arabidopsis thaliana and tobaccos
  • Application of soybean gene promoters pRPS28 and pRPS28-I in soybeans, arabidopsis thaliana and tobaccos

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Embodiment

[0062] one, RPS28 genes and EIF1 gene screening

[0063] The soybean variety Jidou 17 was selected as the material. After sowing, the slow-growing rhizobia USDA110 was inoculated when the true leaves unfolded (8 days after sowing). At 20, 25 and 30 days, samples of root, nodule, hypocotyl, cotyledon, epicotyl, true leaf, true leaf node, compound leaf, internode, petiole and terminal bud were selected respectively. The RNA-seq data is screened, and RNA is extracted for RNA sequencing. The RNA extraction and sequencing methods can use the existing technology, which is not the invention of the present invention, so it will not be repeated. and filtered according to the following criteria:

[0064] 1. According to the soybean RNA-seq data in this experiment, analyze the genes that are expressed in each period and in each tissue site, and sort the top 20 soybean ubiquitous genes as candidate genes according to their FPKM values ​​from large to small.

[0065] 2. Plot the RNA ...

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Abstract

The invention belongs to the technical field of plant genetic engineering, and particularly relates to application of soybean gene promoters pRPS28 and pRPS28-I in soybeans, arabidopsis thaliana and tobaccos. According to the invention, a gene RPS28 and a gene EIF1 are cloned from soybeans; a promoter pRPS28 and a promoter pRPS28-I are obtained through a CTAB method, then a soybean stable transformation vector is constructed, expression of GUS protein driven by the pRPS28 and the pRPS28-I in all stages of soybean genetic transformation is achieved, it is found that the GUS driven by the pRPS28 and the pRPS28-I is expressed in cotyledons, radicles, germs, true leaves, compound leaves, buds, petioles, internodes, roots and root nodules, it is indicated that the pRPS28 and the pRPS28-I are pan-expression promoters, and the activity of the expression product of the GUS protein in the soybean is measured. In addition, the transformation of the GUS protein driven by the pRPS28 and the pRPS28-I in the transformation of the arabidopsis thaliana and the tobacco is also realized.

Description

technical field [0001] The invention belongs to the technical field of plant genetic engineering, in particular to a soybean gene promoter pRPS28 and pRPS28-I Application in soybean, Arabidopsis and tobacco. Background technique [0002] Promoters are an important part of regulating plant gene expression. According to the source, promoters can be divided into exogenous promoters and endogenous promoters. Exogenous promoters can heterologously regulate gene expression, but transgenic safety problems often arise when preparing transgenic materials, especially commonly used promoters derived from viruses, such as the tobacco mosaic virus 35S promoter. In addition, transgene silencing may occur when the same promoter is used in the same transgenic event. Therefore, endogenous promoters are more favored promoters for the preparation of transgenic materials. Promoters are classified into constitutive promoters, tissue-specific promoters, and inducible promoters according to ...

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Application Information

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IPC IPC(8): C12N15/113C12N15/11C12N15/84A01H5/00A01H6/54A01H6/20A01H6/82
CPCC07K14/415C12N15/8223
Inventor 王学路王海娇彭亚齐
Owner HENAN UNIVERSITY
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