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

Sorghum 14-3-3 protein GF146 gene, recombinant vector thereof and expression method

A technology for recombining vectors and genes, applied in botany equipment and methods, biochemical equipment and methods, genetic engineering, etc., can solve problems that have not been reported on 14-3-3 protein gene research

Inactive Publication Date: 2019-06-28
GUIZHOU UNIV
View PDF6 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But so far, there is no research report on 14-3-3 protein gene in sorghum at home and abroad

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
  • Sorghum 14-3-3 protein GF146 gene, recombinant vector thereof and expression method
  • Sorghum 14-3-3 protein GF146 gene, recombinant vector thereof and expression method
  • Sorghum 14-3-3 protein GF146 gene, recombinant vector thereof and expression method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1: Acquisition and Analysis of Sorghum 14-3-3 Protein GF14b Gene

[0033] According to the reported protein sequence of the rice GF14b gene (gene number LOC_Os01g43540), blastP searched the sorghum genome database (https: / / phytozome) to find the sorghum homologous gene GF14b (gene number Sb03g028430). Search the NCBI database with the GF14b protein, download the 14-3-3 genes in other species, and use the MEGA 7.0 software to construct an unrooted evolutionary tree with Neighbor-Joining (NJ) (bootstrap=1000). figure 1 It can be seen that GF14b and sugarcane 14-3-3 genes are clustered into one branch and have the closest relationship.

Embodiment 2

[0034] Example 2: Construction and Identification of Sorghum 14-3-3 Protein GF14b Gene Recombination Vector

[0035] 1. Extract sorghum RNA and reverse transcribe cDNA

[0036] The sorghum BTx623 material was taken, the total RNA at the seedling stage was extracted with an RNA extraction kit (Tiangen Biochemical Technology (Beijing) Co., Ltd.), and cDNA was obtained by reverse transcription with a reverse transcription kit (Promega).

[0037] 2. Using cDNA as a template to amplify the GF14b gene;

[0038] Primers were designed to amplify GF14b gene using cDNA as template.

[0039] Primers are as follows:

[0040] Upstream primer: GF14b-F: GC GAATTC The underline of ATGGCATCAGCAGAGCTTT is the restriction site of BamHI;

[0041] Downstream primer: GF14b-R: CG CTCGAG TTACTGCCCCTCGCTCGA is underlined as the XhoI restriction site;

[0042] The upstream and downstream primers are shown in SEQ ID NO.3 and 4 respectively.

[0043] The PCR amplification system used for gene amp...

Embodiment 3

[0048] Example 3: Induced expression of GF14b protein

[0049] 1. Obtain the recombinant prokaryotic expression strain of GF14b

[0050] The single clone successfully sequenced in Example 2 was selected and inoculated into 50ug / mL kanamycin liquid medium, cultured overnight at 37°C and 200rpm, and the pET-28a -The GF14b recombinant expression vector was extracted, and the recombinant expression vector plasmid was taken to transform Escherichia coli expression strains BL21(DE3), JM109(DE3), Rosetta(DE3), Tuner(DE3), BL21(DE3)pLysS to detect the expression of GF14b protein.

[0051] 2. Cultivate the activated strain overnight

[0052] The above-mentioned recombinant prokaryotic expression strains were activated by culturing overnight. For example, transfer BL21(DE3), JM109(DE3), Tuner(DE3) strains to 50ug / mL kanamycin liquid medium, transfer Rosetta(DE3), BL21(DE3)pLysS strains to 50ug / mL In mL kanamycin+50ug / mL chloramphenicol liquid medium, cultivate the activated strain ov...

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 sorghum 14-3-3 protein GF14b gene. The nucleotide sequence of the gene is shown in SEQ ID NO.1. The invention further discloses a protein encoded by the sorghum 14-3-3 protein GF14b gene, and the amino acid sequence is shown in SEQ ID NO.2. The invention further discloses a recombinant vector containing the sorghum 14-3-3 protein GF14b gene and an expression method. Accordingly, by means of a rice GF14b gene protein sequence, a sorghum homologous gene GF14b is obtained from a sorghum genome database, and the gene is 786 BP in total length; according to the gene, a prokaryotic expression vector is constructed and purified through a protein purification system. The result lays a foundation for further studying the protein crystal structure and biological characteristics, and then the basis is provided for improving the stress resistance of sorghum through a gene editing method.

Description

technical field [0001] The invention relates to a sorghum 14-3-3 protein GF14b gene, a recombinant vector and an expression method thereof, belonging to the field of biotechnology. Background technique [0002] Sorghum (Sorghum bicolor (L.) Moench), also known as water millet and chestnut, is an important cereal crop with strong resistance to adversity. It is an important raw material, and it is also a kind of animal husbandry crop that has been widely used in recent years. Sorghum has a long history of cultivation in my country, and it was not cultivated on a large scale before the founding of the People's Republic of China. It was not until the 1970s, with the development of economy and society, that my country gradually paid attention to and cultivated sorghum on a large scale. Sorghum is a short-day C4 plant. Compared with other energy crops, sorghum has more obvious photosynthetic efficiency and higher yield advantages, so it is known as a "high-energy crop". In recen...

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): C12N15/29C07K14/415C12N15/70
Inventor 谢鑫陈俊李向阳蒋君梅任明见孙涛屈志广冯泽锐黄磊王营锴陈美晴方远鹏
Owner GUIZHOU UNIV
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