Eureka AIR delivers breakthrough ideas for toughest innovation challenges, trusted by R&D personnel around the world.

Application of rice transcription factor Os02g49250 gene

A rice transcription factor and gene technology, applied in the field of genetic engineering, can solve problems such as the weakening of hybrid breeding advantages

Inactive Publication Date: 2014-12-17
INST OF CROP SCI CHINESE ACAD OF AGRI SCI
View PDF2 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The current research on increasing rice production is more dependent on limited rice germplasm resources, the advantages of traditional hybrid breeding are gradually weakening, and rice transgenic technology may explore the potential of further increasing rice production

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
  • Application of rice transcription factor Os02g49250 gene
  • Application of rice transcription factor Os02g49250 gene
  • Application of rice transcription factor Os02g49250 gene

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] The separation of embodiment 1Os02g49250 gene and the construction of plant expression vector

[0036]Find the Os02g49250 gene in the plant transcription factor database, design PCR amplification primers according to its sequence (F1: 5'-CAAAAAAGCAGGCTTCATGGAGGAGGAGAAGGAGGAGA-3' and reverse primer R1: 5'-CAAGAAAGCTGGGTC GACCGACATGAACGCGA AATC- 3'). The complete sequence of Os02g49250 was obtained by PCR using the total cDNA of wild-type Nipponbare rice as a template, and its nucleotide sequence is shown in SEQ ID No.1.

[0037] Perform PCR according to the PrimeSTAR polymerase amplification system and reaction procedures. This process includes two rounds of PCR. The primers of the first round of PCR use the gene primers (F1 and R1) with a partial adapter attB linker, while the template of the second round uses the PCR product of the first round, and the primers use the complete adapter attB Primers (attB5'adaptor: 5'-GTGGGGACAAGTTTGTACAAAAAAGCAGGCTTC-3', attB3'adaptor...

Embodiment 2

[0038] The acquisition of embodiment 2 transgenic rice plants

[0039] Take the mature seeds of rice 'kitaake', shell them manually or mechanically, select the plump, smooth and spot-free seeds and inoculate them on the induction medium for induction culture after being sterilized. Rice callus with good appearance and good growth was selected as the recipient material, and ubi:VP64-Os02g49250 was transferred into the rice callus by the Agrobacterium-mediated method, and acetosyringone containing 100 μM and O.D. value of 0.7 were used. The AAM transformation liquid of Agrobacterium was transformed, and the callus soaked in the transformation liquid was placed on the co-culture medium for co-cultivation, cultured in the dark at 25°C for 3 days, then placed on the screening medium for about 30 days, and subcultured every 10 days. Then, the screened out resistant calli were transferred to the differentiation medium for differentiation for about 20 days, and subcultured every 10 da...

Embodiment 3

[0044] Identification of embodiment 3 transgenic positive strains

[0045] In order to detect the overexpression of ubi: VP64-Os02g49250 gene in T2 transgenic rice, the VP64 antibody was used to identify it at the protein level, after SDS-PAGE protein electrophoresis→immunoblotting→immunofluorescence reaction, the results of Western Blot identification showed that The transgenic plants had the target protein, while the wild type had no bands ( image 3 ).

[0046] The specific Western Blot experiment process is as follows:

[0047] Put an appropriate amount of sample into liquid nitrogen and freeze it, grind it into powder, add an appropriate amount of loading buffer, mix well, and centrifuge at 12000rpm for 10 minutes; take 5μl of the supernatant and add the sample, electrophoresis at 90V for SDS-PAGE for 30 minutes, and electrophoresis at 120V for 60-90 When the bromophenol blue reaches the bottom of the gel, the electrophoresis can be stopped; after electrophoresis, the m...

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 relates to an application of a rice (oryza sativa l.) transcription factor Os02g49250 gene. According to the application, transcription factor activated sequence motifs VP64 is utilized to be fused with the rice transcription factor Os02g49250 gene to construct a combined type transcription factor, and the combined type transcription factor is transformed into crops such as rice, so as to improve rice grain traits; for example, the rice grain length and width are increased. The application has an important theoretical value in illuminating regulation and control of a seed development mechanism in detail and also can improve the rice grain type through a genetically modified method, so that the application also has important significance in production practices.

Description

technical field [0001] The invention relates to the field of genetic engineering, in particular to the application of rice transcription factor Os02g49250 gene. Background technique [0002] Rice (Oryza sativa L.) is one of the three most important food crops in my country and the world, the staple food of more than half of the world's population, and an important model plant for functional gene research. Related genetics and molecular biology studies have been paid much attention by researchers, and the regulation of transcription level is an important way of gene expression regulation. The current research on increasing rice yield is more dependent on limited rice germplasm resources, the advantages of traditional hybrid breeding are gradually weakening, and rice transgenic technology may explore the potential of further increasing rice yield. [0003] In the plant kingdom, plants that can form seeds account for more than two-thirds of the total number of plants. As impor...

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): C07K19/00C12N15/62C12N15/82C12N1/21C12N15/11A01H5/00
Inventor 赵涛张春雨刘斌李宏宇刘军林辰涛
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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
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