Plant drought induced type synthesis promoter SP15 and application thereof

An inducible, promoter technology, applied in the fields of biotechnology and genetic engineering, can solve the problem that natural promoters cannot finely regulate genes

Active Publication Date: 2019-06-04
JILIN AGRICULTURAL UNIV
View PDF5 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to solve the problem that the natural promoter cannot finely regulate the gene, and provide a plant drought-inducible promoter SP15

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Plant drought induced type synthesis promoter SP15 and application thereof
  • Plant drought induced type synthesis promoter SP15 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] Example 1 Design of plant drought-inducible promoter SP15

[0016] By combining the soybean stress transcriptome sequencing data with the existing soybean stress expression profile data in Genbank, 63 target genes regulated by drought were screened out, and then they were listed for cluster module classification; after obtaining 4 clusters, they were identified using motifs The software identifies the motifs of each cluster separately, and finally obtains 46 shared expression elements of promoters related to stress regulation; among them, G-box, ABRE, Root specific motif, MYB1, MYB4, and GCC-box are selected as the basic cis Elements, with 2 copies of G-box elements, 1 copy of ABRE elements, 2 copies of Rootspecific motifs, 2 copies of MYB1 elements, 2 copies of MYB4 elements, 2 copies of GCC-box elements, 2 copies Two copies of the Root specific motif constitute a tandem unit, with a specific sequence interval of 10 bases between each two elements, and the above-design...

Embodiment 2

[0017] Example 2 Construction of promoter SP15 expression vector

[0018] The sequence of SP15 (SEQ ID No.1) was synthesized into the vector pUC57 by artificial synthesis method, and enzyme cutting sites were added at both ends of the sequence Sac l and Nco I; the pUC57-SP15 vector and the pCAMBIA3301 plasmid were used respectively Sac l and Nco I carried out double enzyme digestion, after recovering the enzyme digestion products respectively, through connection, transformation and identification, the expression vector pCAMBIA3301-SP15 was obtained ( figure 1 ); the vector pCAMBIA3301-SP15 was transformed into Agrobacterium rhizogenes K599 for infecting soybean cotyledonary nodes.

Embodiment 3

[0019] Example 3 Obtaining of Transgenic SP15 Gene Hairy Roots of Soybean

[0020] Using the established soybean hairy root induction system, Agrobacterium rhizogenes K599 containing the pCAMBIA3301-SP15 carrier was used to infect soybean cotyledon nodes. After induction and identification, transgenic positive hairy roots were obtained; drought stress treatment was carried out; the specific method was as follows :

[0021] (1) After mixing sodium hypochlorite and concentrated hydrochloric acid 1:1, sterilize soybean seeds with chlorine gas overnight;

[0022] (2) Wash the soybeans with sterile water and plant them in high-temperature and high-pressure sterilized vermiculite, and moisturize with 1 / 4 Hoagland's solution;

[0023] (3) Cultivate for about 6 days in an artificial climate chamber at 25°C, under light for 16 hours;

[0024] (4) Agrobacterium rhizogenes K599 containing the recombinant plasmid pCAMBIA3301-SP15 was cultured overnight in an incubator at 28°C;

[0025]...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention discloses a plant drought induced type synthesis promoter SP15. The nucleotide sequence of the plant drought induced type synthesis promoter SP15 is shown as SEQID No.1. A plant expression vector is formed by that a nucleotide sequence as shown in the SEQID No.1 is inserted to a vector plasmid. The invention further relates to an application of the promoter SP15 to breeding of drought resisting genetically-modified plant, and belongs to an artificially synthesized promoter. The expression situation of the SP15 promoter driving report genes GUS identified in a soybean hairy root transformation system indicates that the SP15 promoter can be induced by drought stress, and the expression intensity is equivalent to a CaMV 35s promoter.

Description

technical field [0001] The invention belongs to the field of biotechnology and genetic engineering, and specifically relates to a plant drought-inducible promoter SP15 and its application. Background technique [0002] Drought affects plants in many ways and is an extremely complex process. Drought will not only affect the growth and development of plants, but also break the balance of physiological and biochemical metabolism of plants, which will eventually lead to a decrease in yield and quality. A large number of scientific studies have shown that plants can solve the adverse reactions caused by drought by regulating the expression differences of key genes in drought-stressed environments. Promoter is a molecular switch to achieve differential expression of key genes, which can control the transcription initiation, expression intensity and expression site of genes, etc., and can be used as a powerful tool for genetic engineering breeding. [0003] A promoter is a DNA co...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12N15/82A01H5/00
Inventor 李海燕李晓薇王法微王南刘伟灿周永刚董园园刘秀明姚娜刘欣高延妍
Owner JILIN AGRICULTURAL UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap