Supercharge Your Innovation With Domain-Expert AI Agents!

Arcelin-5 promoter and uses thereof

a technology of arcelin-5 and promoter, applied in the field of plant genetics, can solve the problems of insufficient accumulation of engineered protein, insufficient gene expression of plant and seed protein, and inability to express genes in plants and seeds,

Inactive Publication Date: 2010-09-30
WANG QI +3
View PDF19 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the protein content of plants and seeds is often incomplete.
Despite the availability of many molecular tools, the genetic modification of plants and seeds is often constrained by an insufficient accumulation of the engineered protein.
This early step is often rate-limiting relative to subsequent stages of protein production.
However, none of these promoters have been reported to demonstrate comparable activity in transgenic soybean plants.
Thus although expression of an Arcelin species was reported, the effectiveness of such Arcelin promoters in crops such as maize and soybean remain totally unknown.

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
  • Arcelin-5 promoter and uses thereof
  • Arcelin-5 promoter and uses thereof
  • Arcelin-5 promoter and uses thereof

Examples

Experimental program
Comparison scheme
Effect test

example 1

Production of Arcelin Promoters

[0170]Seeds of the P. vulgaris exotic genotype G02771 containing the Arcelin-5 seed protein are obtained from the USDA seed stock center (USDA, ARS, Washington State University, Regional Plant Introduction Station, 59 Johnson Hall, P.O. Box 646402, Pullman, Wash., USA 99164-6402). A PCR approach is designed and optimized to obtain fragments from the Arcelin-5 nucleic acid sequence. The following six fragments from Arcelin-5 are obtained:

[0171]The promoter region (1.8 Kb); the promoter region upstream of the transcription initiation site (TATA box) (1.7 Kb); the 3′UTR region (terminator) (1.3 Kb); the promoter region with a substituted 5′UTR (dSSU soybean 5′UTR) (1.8 Kb); the genomic clone (4 Kb); and the coding region.

[0172]Exotic genotypes containing other Arcelin promoters (e.g., Arcelin-3 (PI 41683), Arcelin-4 (PI 417775), are similarly obtained. The Arcelin-3 and Arcelin-4 promoters are cloned using the PCR approach described above. A sequence alig...

example 2

Transformation of Soybean Cotyledons

[0174]Soybean cotyledons are transiently transformed with pMON58100, 13773, 55524, 55527, 55538, and 55539 (see Table 2 for description of each) by particle bombardment. Briefly, seeds from Asgrow A3244 soybean plants are harvested 25-28 days after flowering. Seeds are osmotically treated overnight at 25° C. in the dark on Gamborg's Media (G5893, Sigma Aldrich Company, St. Louis, Mo.) supplemented with 50 mM glutamine, 111 mM maltose, 125 mM raffinose, 125 mM mannitol, and 3 g / L purified agar, pH 5.6. The resulting half seeds are bombarded with 2 μg / L of the Arcelin and 7Sα′ promoter constructs. A separate e35S driven luciferase construct is included in a 1:1 molar ratio with each of the promoter constructs as an internal control. Bombarded tissues are incubated for 48 hours at 25 C.

example 3

Analysis of Arcelin Promoter Activity

[0175]Bombarded tissues are analyzed for expression of GUS and luciferase activity. Protein is extracted from six bombarded soybean cotyledons using 1 ml extraction buffer containing 0.1 M potassium phosphate (pH 7.8), 10 mM DTT, 1 mM EDTA, 5% glycerol, and protease inhibitor cocktail (1 tablet / 50 ml; #1697498, Roche Diagnostics Corporation, Indianapolis, Ind.). Protein extract is prepared in 100 μL aliquots and tested for luciferase activity according to manufacturer protocol (Steady-Glo™, #E2510, Promega Corporation, Madison, Wis.). GUS activity is measured using 50 μl aliquots following a standard procedure with minor modification (Maliga, et al., 1995, :Methods in Plant Molecular Biology, A Laboratory Course Manual,” Cold Spring Harbor Laboratory Press, page 29). GUS activity is normalized based on the luciferase activity of the internal control. Results of three independent experiments (performed in duplicate) are provided in FIG. 2. The res...

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 present invention discloses Arcelin promoters capable of transcribing a heterologous nucleic acid sequence at high levels in plants. The promoters are particularly suited for use in soybean plants and plant cells. Methods of modifying, producing, and using the promoters are also disclosed. The invention further discloses compositions, transformed host cells, transgenic plants, and seeds containing the high-expression Arcelin promoters, and methods for preparing and using the same.

Description

CROSS REFERENCE TO RELATED APPLICATION[0001]This application claims priority to U.S. Provisional Application Serial No. 60 / 255,879, filed Dec. 18, 2000, and having the title “Arecelin 5 Promoter and Uses Thereof”, which is herein incorporated by reference in its entirety.FIELD OF THE INVENTION[0002]The present invention generally relates to the field of plant genetics. More specifically, the present invention relates to gene expression in plants. The invention provides promoters capable of transcribing a heterologous nucleic acid sequence at a high level in plants, and methods of modifying, producing, and using the same. The invention also provides transformed host cells, transgenic plants, and seeds containing the high-expression promoters, and methods for preparing and using the same.BACKGROUND OF THE INVENTION[0003]Plants and seeds provide an important source of dietary protein for humans and livestock. However, the protein content of plants and seeds is often incomplete. For exa...

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(United States)
IPC IPC(8): A01H5/00C12N5/10C07K14/415C12N15/29C12N15/82
CPCC07K14/415C12N15/8234C12N15/8216A61P1/00
Inventor WANG, QIDUBOIS, PATRICELIANG, JIHONGOULMASSOV, TIM
Owner WANG QI
Features
  • R&D
  • Intellectual Property
  • Life Sciences
  • Materials
  • Tech Scout
Why Patsnap Eureka
  • Unparalleled Data Quality
  • Higher Quality Content
  • 60% Fewer Hallucinations
Social media
Patsnap Eureka Blog
Learn More

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

© 2025 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com