Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Improved protein production and storage in plants

A protein, heterologous protein technology, applied in the field of plant genetics, can solve the problems of no teaching, reduction, etc.

Inactive Publication Date: 2011-07-27
DONALD DANFORTH PLANT SCI CENT +1
View PDF19 Cites 3 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Wu does not teach reducing expression of β-conglycinin or glycinin or expressing exogenous proteins

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
  • Improved protein production and storage in plants
  • Improved protein production and storage in plants
  • Improved protein production and storage in plants

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0210] Generation of storage protein knockdown line "SP-"

[0211] Transformation protocol using gene gun (Parrott et al., 2004, "Transgenic soybean," In: J.E.Specht and H.R.Boerma (eds). Soybeans: Improvement, Production, and Uses, 3rd Edition - Agronomy Monograph No. 16.ASA-CSA- SSSA, Madison, WI.pp 265-302), will be based on figure 2 RNAi constructs designed to suppress storage protein content in seeds were transferred to soybean. Specific simultaneous inhibition of endogenous soybean storage protein and FAD2-1ω-6 was generated by inversion of the sequence specific for this type of open reading frame, which flanks the intron, under the control of the glycinin promoter and 3' terminator RNAi cassette for fatty acid desaturase. The 331 bp region (SEQ ID NO: 55) of the glycinin AlbB2 gene was placed next to the 128 bp region (SEQ ID NO: 57) of the FAD2-1a gene. This 459 bp heterologous DNA was then placed around the intron as an inverted repeat. Introns of synthetic origi...

Embodiment 2

[0215] Protein and oil content in SP-

[0216] The SP-lines were regenerated into soybean plants. The resulting plant growth and fruit set were not significantly different from the control. TO seeds were broken to determine the phenotype, SP-seeds were regrowth and selected two more times to generate a homozygous population. The SP-phenotype was stable in each subsequent generation with no detectable α / α' and β conglycinin subunits and greatly reduced glycinin levels. Oleic acid levels were higher than 94% in SP-seeds, indicating that there was also FAD2 selection marker knockdown.

[0217] The average dry size and weight of 146 mg of dormant seeds of SP- grown in the greenhouse was similar to that of the wild-type (WT) variety "Jack" grown in the greenhouse (mean 163 mg). The total protein and oil content of SP- (40.2%, 19.1%) was similar to that of WT variety "Jack" (37.5%, 20.5%). Thus, this assay demonstrated that knockdown of proteins corresponding to most of the tota...

Embodiment 3

[0219] Electron microscopy and immunogold immunocytochemistry

[0220] Tissue samples were freeze-fixed using a Balzer's high pressure apparatus (Bal-Tech, Principality of Liechtenstein), cryo-displaced with acetone / OsO4, and embedded in epon plastic. Ultrathin sections were stained with saturated uranyl acetate aqueous solution and lead citrate (33 mg / ml), and then observed. Immunocytochemical analysis was then performed. Freeze-fixed parallel samples were then processed by freeze-substitution in the absence of fixative. The displaced samples were transferred to Lowicryl HM-20 resin (which was polymerized by UV light irradiation). Thin sections were labeled with anti-GFP MAb (Clontech) or rabbit polyclonal anti-glycinin previously generated by our laboratory. Sections were indirectly labeled with anti-IgG (rabbit or mouse)-10 nm colloidal gold (Sigma) and then 5% uranyl acetate to increase contrast prior to EM observation. All TEMs were performed with a LEO 912AB microsco...

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

PropertyMeasurementUnit
molecular weightaaaaaaaaaa
molecular weightaaaaaaaaaa
Login to View More

Abstract

A transgenic dicotyledonous plant has a deficiency of one or more plant seed storage proteins, further has a transgenic polynucleotide construct comprising an open reading frame operably linked to a storage protein promoter and an ER signal sequence. The polynucleotide construct encodes a protein product that can accumulate at high levels in the seed. Also provided are methods of producing a heterologous protein in a plant seed.

Description

[0001] Cross Reference Related Applications [0002] This application claims priority to US Provisional Application Serial No. 61 / 076,616, filed June 28, 2008, which is hereby incorporated by reference in its entirety. [0003] Statement Regarding Federally Sponsored Research or Development [0004] This invention was supported in part by the U.S. Government under Special Research Grant #2006-06103 awarded by USDA and administered by the University of Missouri, Columbia, and by an intramural research grant from the USDA Agricultural Research Service (Project #3622-210000-025- 00). The US Government may have certain rights in this invention. [0005] Name and participants of the joint research agreement [0006] This invention arose out of a joint research agreement between the Donald Danforth Plant Science Center and the United States Government, represented by the U.S. Department of Agriculture, Agricultural Research Service ("USDA"). Accordingly, the US Government may have...

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): C12N15/11C12N5/14
CPCC12N15/8258C12N15/8257C12N15/8251C12N15/8234C12N9/2434
Inventor E·M·赫曼M·A·施密特
Owner DONALD DANFORTH PLANT SCI CENT
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com