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Application of soybean gene promoters peif1 and peif1-i in soybean, Arabidopsis and tobacco

A promoter and Arabidopsis technology, applied in the fields of application, genetic engineering, plant gene improvement, etc., can solve the problems of imperfect screening conditions for constitutive promoters, gene silencing, and no expression related to soybean nodule development

Active Publication Date: 2022-06-21
HENAN UNIVERSITY
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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 K. McHale, John J. Finer. Isolation and characterization of “GmScream” promoters that regulate highly expressing soybean (Glycine maxMerr.) genes[J]. Plant Science, 2015, 241.) According to soybean strains The RNA sequencing data of 31 libraries of Williams82 (WS82) screened 10 expression genes of possible constitutive promoters, but none of them involved a continuous process of soybean growth and development and expression in root nodule development
At the same time, when multiple genes are transferred in the same plant, it is easy to cause gene silencing if driven by the same promoter

Method used

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  • Application of soybean gene promoters peif1 and peif1-i in soybean, Arabidopsis and tobacco
  • Application of soybean gene promoters peif1 and peif1-i in soybean, Arabidopsis and tobacco
  • Application of soybean gene promoters peif1 and peif1-i in soybean, Arabidopsis and tobacco

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Experimental program
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Embodiment

[0063] one, RPS28 genes and EIF1 gene screening

[0064] 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:

[0065] 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.

[0066] 2. Plot the RNA ...

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Abstract

The invention belongs to the technical field of plant genetic engineering, in particular to a soybean gene promoter pEIF1 and pEIF1‑I Application in soybean, Arabidopsis and tobacco. The present invention clones the gene from soybean RPS28 and genes EIF1 ; Get the promoter by CTAB method pEIF1 and promoter pEIF1‑I , and then construct a soybean stable transformation vector, and realize pEIF1 and pEIF1‑I The expression of the driven GUS protein at each stage of soybean genetic transformation, found that pEIF1 , pEIF1‑I The driven GUS is expressed in cotyledons, radicles, germs, true leaves, compound leaves, buds, petioles, internodes, roots and nodules, indicating that pEIF1 and pEIF1- I It is a ubiquitous expression promoter, and the activity of GUS protein expression product in soybean is determined. In addition, it also implements pEIF1 and pEIF1‑I Driving transformation of GUS proteins in Arabidopsis and tobacco transformation.

Description

technical field [0001] The invention belongs to the technical field of plant genetic engineering, in particular to a soybean gene promoter pEIF1 and pEIF1-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. According to the expression characteristics, promoters are divided into constitutive promoters, tissue-specific promoter...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/113C12N15/11C12N15/84A01H5/00A01H6/54A01H6/20A01H6/82
CPCC07K14/415C12N15/8223C12N15/8216
Inventor 王学路王海娇彭亚齐
Owner HENAN UNIVERSITY
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