Plant drought-induced artificial synthesis promoter SP2 and application thereof

A promoter, inducible technology, applied in the field of genetic engineering, can solve the problems of low expression activity and low specificity

Inactive Publication Date: 2017-09-15
JILIN AGRICULTURAL UNIV
View PDF0 Cites 8 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the activities of the natural promoters currently isolated and iden

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 artificial synthesis promoter SP2 and application thereof
  • Plant drought-induced artificial synthesis promoter SP2 and application thereof
  • Plant drought-induced artificial synthesis promoter SP2 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0017] Example 1. Design of plant stress-inducible promoter SP2

[0018] By combining the soybean stress transcriptome sequencing data with the existing soybean stress expression profile data in Genbank, 63 target genes that are regulated by drought were screened out, and then they were listed for cluster module classification. After obtaining 4 clusters, motif identification software was used to identify each cluster separately, and finally 46 promoter shared expression elements related to stress regulation were obtained. Select G-box, ABRE, ABF, SORLIP1, CCA1, E2F as the basic cis-elements. Each element is repeated 4 times in tandem. The two elements are separated by a random sequence of 7-9 bases. Synthetic Promoter 2 is constructed by connecting with 35S core promoter Core CaMV 35S, which is called SP2 for short, and its nucleotide sequence is SEQ ID No. 1 sequence.

Example Embodiment

[0019] Example 2. Construction of promoter SP2 expression vector

[0020] Synthesize the SP2 (SEQ ID No.1) sequence into the vector pUC57 by artificial synthesis, and add restriction sites at both ends of the sequence BamH l and Hind III. The pUC57-SP2 vector and pBI121 plasmid were used BamH l and Hind III Carry out double enzyme digestion, and recover the digested products separately, and obtain the expression vector pBI121-SP2 (such as figure 1 ). The vector pBI121-SP2 was transformed into Agrobacterium EHA105 for infecting tobacco.

Example Embodiment

[0021] Example 3. Obtaining SP2 transgenic tobacco

[0022] Follow the leaf disc transformation method (Horsch et al. 1988). The Agrobacterium liquid is diluted with sterilized water to 106-107cells / mL (generally diluted about 30 times). Take sterile tobacco leaves, remove the main veins and cut the leaves into square shapes, and then immerse them in the diluted bacterial solution for 8-10 minutes.

[0023] 1) Co-cultivation

[0024] After taking out the leaf explants, blot the bacteria liquid on sterilized filter paper, place the leaf surface down on the surface of the MS medium, and cultivate in the dark at 24°C-25°C for 2 days.

[0025] 2) Select training

[0026] The co-cultured leaves were transferred to selective differentiation medium (containing 250 mg / L cephalosporin; 100 mg / L kanamycin), 28°C, 16 hours of light per day, and the medium was changed every 15-20 days.

[0027] 3) Rooting culture

[0028] When the differentiated resistant buds grow to about 1 cm, cut them off (pref...

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 soybean-induced promoter SP2 and belongs to the construction of artificial synthesis promoters. The soybean-induced promoter SP2 is formed by taking G-box, ABRE, ABF, SORLIP1, CCA1 and E2F as basic cis elements, repeatedly connecting all the elements in series for four times, spacing every two elements through a random sequence of nine bases and then connecting a design fragment and a 35S key promoter Core-CaMV-35S, and is called SP2. The identification on the expression situation of an SP2 promoter-driven reporter gene GUS in transgenic tobaccos shows that the promoter SP2 can be induced by drought, salt, saline, ABA and low-temperature stress and is extremely sensitive to drought stress. The soybean-induced promoter SP2 disclosed by the invention can be directly applied to crop transgenic resistance seed breeding.

Description

technical field [0001] The invention belongs to the field of genetic engineering, in particular to a plant drought-inducible promoter SP2 and its application in transgenic plants. technical background [0002] A promoter regulates the expression of a gene by binding to one or more transcription factors. Each promoter has its specific sequence structure and binding site. The main structure of the promoter includes the core promoter sequence and upstream promoter elements, which can guide the transcription of nearby DNA segments. Promoters play a very important role in the initiation and regulation of gene transcription, and are important building blocks in genetic engineering. In recent years, with the development of synthetic biology, artificially synthesized functional elements have shown great application potential in the field of metabolic engineering. The metabolic flux of cells is mainly regulated by the transcriptional level, and the promoter is an important regulato...

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
IPC IPC(8): C12N15/113C12N15/82C12N15/62A01H5/00
CPCC07K14/415C12N15/8273
Inventor 李海燕李晓薇王法微刘伟灿王南董园园刘秀明姚娜刘欣肖洪庆戢舒涵
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