Unlock instant, AI-driven research and patent intelligence for your innovation.

Soybean salt-induced artificially synthesized promoter AP2 and application thereof

An artificial synthesis and promoter technology, applied in the field of promoters, can solve problems such as low specificity and low expression activity

Active Publication Date: 2019-01-11
SHANDONG UNIV
View PDF3 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the activity of the isolated and identified natural promoters is limited to some extent, such as low expression activity and low specificity. Therefore, it is of great importance to develop and utilize highly active synthetic promoters to achieve precise regulation of target genes. scientific and practical significance

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
  • Soybean salt-induced artificially synthesized promoter AP2 and application thereof
  • Soybean salt-induced artificially synthesized promoter AP2 and application thereof
  • Soybean salt-induced artificially synthesized promoter AP2 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] The design of embodiment 1 soybean salt-inducible artificial synthetic promoter

[0016] The applicant's previous research found that the 300bp (P4) promoter region of the GmST1 gene of Shengdou No. 9 was the basic promoter, and the genomic DNA of Shengdou No. 9 was extracted by CTAB method, and the extracted genomic DNA was diluted to 100ug / ul for amplification Promoter template. Using the high-fidelity enzyme HIFI (Takara) to amplify the target fragment and sequence it, and simultaneously synthesize six tandem repeats of GT1GMSCAM4 cis-element nucleic acid sequences that respond to salt stress, and connect the two sequences to construct a synthetic promoter, named It is a soybean salt-inducible artificially synthesized promoter AP2, and the nucleotide sequence of the promoter is shown in SEQ ID No.1.

Embodiment 2

[0017] Example 2 Using Arabidopsis protoplast transient transformation technology to verify that the salt-inducible artificial promoter AP2 in soybean is induced by salt.

[0018] 2.1 Construction of pGmST1::BM2::P4::pGreenII-0800LUC vector

[0019] 2.1.1 Cloning of the AP2 promoter fragment containing restriction sites

[0020] The dual fluorescent reporter vector pGmST1::BM2::P4::pGreenII-0800LUC of the soybean salt-inducible artificial promoter AP2 was constructed by double enzyme digestion-ligation method. Using the restriction site analysis on Primer5 AP2 and comparing it with the multiple cloning site on pGreen II-0800LUC, the available restriction sites were kpnI and SpeI.

[0021] PCR amplification was performed using primers containing restriction enzyme sites in AP2.

[0022] The reaction system for high-fidelity enzyme HIFI amplification is as follows (20 μl system):

[0023]

[0024]

[0025] The amplification conditions are as follows:

[0026]

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 salt-induced artificially synthesized promoter AP2, being characterized in that the promoter region of GmST1 gene in Soybean Shengdou 9 is 300bp (P4) and six tandem repeats of GT1GMSCAM4 cis-element are ligated. The nucleotide sequence of the promoter is shown as SEQ ID No. 1. The promoter region is cloned into the GT1GMSCAM4 cis-element, which is responsive to salt stress. As prove by that relative luciferase activity experiment of the protoplast transformed by the promote LUC plasmid, the promoter AP2 of the invention can quickly and efficiently respond to the high-salt induction, which indicates that the promoter is sensitive to salt stress, and can be apply to cultivate salt-tolerant transgenic plants and crop transgenic stress-resistant breeding.

Description

technical field [0001] The invention relates to a promoter, in particular to a soybean salt-inducible artificial synthetic promoter AP2 and its application. Background technique [0002] To achieve fine regulation of gene expression, including tissue-specific expression and inducible expression, is how to use molecular switch promoters to regulate the expression of a single gene or multiple genes for specific environmental, physiological and chemical inductions. In recent years, with the development of biosynthesis, artificially synthesized functional elements have played a huge role in the field of metabolic engineering, and promoters are important regulatory elements at the transcription level. Therefore, the fine regulation of genes by artificially synthesized promoters is important in plant growth and development. , tissue characteristics, biotic and abiotic resistance will play an important role. [0003] A cis-acting element is a nucleic acid sequence located upstream...

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): C12N15/113C12N15/82A01H5/00A01H6/20
CPCC07K14/415C12N15/8238C12N15/8273
Inventor 向凤宁孙家旗张文晓吕孟华刘振华李朔
Owner SHANDONG UNIV