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

Application of soybean bHLH transcription factor GmPIF1 gene in promoting isoflavone synthesis

A technology of transcription factors and isoflavones, which is applied in the fields of genetic engineering and molecular biology, can solve the problems that soybean GmPIF1 gene research has not been reported, and achieve the effect of high-efficiency synthesis

Active Publication Date: 2020-09-22
JILIN UNIV
View PDF7 Cites 7 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] So far, the study of soybean GmPIF1 gene has not been reported

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 soybean bHLH transcription factor GmPIF1 gene in promoting isoflavone synthesis
  • Application of soybean bHLH transcription factor GmPIF1 gene in promoting isoflavone synthesis
  • Application of soybean bHLH transcription factor GmPIF1 gene in promoting isoflavone synthesis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029]Example 1: Cloning of the GmPIF1 transcription factor gene and its biological analysis

[0030] A group of genes whose expression levels increase with the development of soybean embryos were screened out from the constructed digital gene expression profiles of soybean Jilin 32 seeds at different stages. These genes are considered to have the potential to regulate the synthesis of soybean isoflavones, and GmPIF1 is one of them. bHLH class of transcription factors. Taking 40 mg of soybean Jilin 32 three compound leaves as a sample, RNAisoReagent (purchased from TaKaRa) was used to extract the total RNA of soybean variety Jilin 32 leaves, and the integrity of the RNA was detected by 1% agarose electrophoresis. Synthesis of cDNA was carried out according to the instructions of Reverse Transcriptase M-MLV (RNase H-).

[0031] According to the CDS sequence of GmPIF1 in NCBI, primer sequences were designed, and the full-length PCR product of CDS of GmPIF1 was obtained by rever...

Embodiment 2

[0042] Example 2: Tissue-specific expression analysis of GmPIF1 gene

[0043] The total RNA of the roots, stems, leaves, flowers, 20-day embryos, 30-day embryos, and 50-day embryos of soybean variety Jilin 32 were extracted and reverse-transcribed into cDNA. The method was the same as in Example 1. The expression of GmPIF1 gene in different soybean tissues was detected by relative fluorescence quantitative PCR (qPCR), according to SYBR○R Premix Ex Taq TM II (purchased from TaKaRa) instructions, using 7500 real-time fluorescence quantitative amplification instrument.

[0044] Using soybean β-tubulin as an internal reference gene, the primer sequence is:

[0045] tubulin-F:5'-GGAAGGCTTTCTTGCATTGGTA-3'

[0046] tubulin-R:5'-AGTGGCATCCTGGTACTGCA-3'

[0047] The quantitative primers for the GmPIF1 gene are:

[0048] GmPIF1-q-F: 5'-AAACCCAAATGGACGACGACTTA-3'

[0049] GmPIF1-q-R: 5'-TGGCCGTTTTGCCATAACAGTTC-3'

[0050] The PCR reaction system is as follows:

[0051]

[0052]...

Embodiment 3

[0055] Example 3: Subcellular Localization of Soybean GmPIF1 Gene

[0056] According to the CDS sequence of GmPIF1, primer sequences with restriction endonucleases BamH1 and Spe1 were designed, and the sequence of GmPIF1 containing restriction sites was obtained through subcloning.

[0057] The primer sequences are:

[0058] GmPIF1-YFP-F: 5'-CGGGATCCATGAATCACTGTGTTCCA-3'

[0059] GmPIF1-YFP-R: 5'-GGACTAGTTCATGTTTCGTCTGACTG-3'

[0060] According to the steps in Example 1, the PCR product obtained by subcloning was connected to the pMD18-T vector, then transferred into Escherichia coli DH5α and sent for sequencing, and the sequenced correct bacterial liquid preservation and extraction plasmid (pT-GmPIF1 / BamH1 / Spe1) were used for future use . The pT-GmPIF1 / BamH1 / Spe1 and the pHB-YFP plant expression vector with the CaMV 35S promoter were double-digested with BamH1 and Spe1 respectively, and the target fragments were gel-cut and recovered, and the recovered GmPIF1 fragment and ...

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
Login to View More

Abstract

The invention relates to application of a soybean bHLH transcription factor GmPIF1 gene in promoting isoflavone synthesis, and belongs to the field of gene engineering and molecular biology. Accordingto the application of the soybean bHLH transcription factor GmPIF1 gene in promoting isoflavone synthesis, the nucleotide sequence of the soybean bHLH transcription factor GmPIF1 gene is shown as SEQID No. 1. The application has the beneficial effects as follows: the GmPIF1 promotes accumulation of isoflavone in soybean hairy roots, a new method is provided for increasing the content of soybeanisoflavone, the soybean isoflavone can be synthesized more efficiently through the bioengineering technology and the gene regulation and control method, and the defects that the artificial cultivationbreeding period is long, hybridization traits are unstable, and screening is difficult are overcome. The transcription factor GmPIF1 gene is transferred into soybean hairy roots, so that the transcription factor GmPIF1 gene is overexpressed in the hairy roots, the content of isoflavone in the hairy roots is increased, and the expression of related enzyme genes in an isoflavone synthetic pathway is regulated.

Description

technical field [0001] The invention relates to the fields of genetic engineering and molecular biology, in particular to the application of a transcription factor gene GmPIF1 mediated by red light and far-red light in soybean to promote the synthesis of isoflavones. Background technique [0002] Soybean (Glycine max) is an important economic crop, which can provide human beings with metabolites such as vegetable oil, vegetable protein and isoflavones. Among them, soybean isoflavones have important commercial value as health products, and can reduce the risk of cardiovascular disease and hyperlipidemia. It has good effects in reducing morbidity, preventing breast disease and osteoporosis, anti-oxidation, alleviating menopausal syndrome, and anti-tumor. In response to market demand, cultivating soybean varieties with high isoflavone quality has attracted extensive attention of researchers. [0003] Soybean isoflavones are secondary metabolites in the phenylalanine metabolic ...

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): C12N15/29C12N15/82C07K14/415A01H5/06A01H6/54
CPCC07K14/415C12N15/8243
Inventor 王庆钰王天亮刘雅婧王英李景文闫帆王硕张鑫生杨旭光
Owner JILIN 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