Transcription factor ERF related to soybean stress, coding gene thereof and application of coding gene
A technology of transcription factors and coding genes, applied in application, genetic engineering, plant genetic improvement, etc., can solve the problems of soybean yield reduction, economic loss, biotic and abiotic stress, etc., and achieve the effect of improving stress resistance
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0024] Embodiment 1: Cloning and sequence analysis of GmERF6 gene
[0025] Soybean RNA extraction and cDNA synthesis: RNAplant plus Reagent (purchased from TIANGEN) was used to extract total RNA from leaves of soybean Jilin 32, and M-MLV reverse transcriptase (RNase H-) (purchased from TaKaRa) was used to perform reverse transcription to synthesize cDNA.
[0026] Design and synthesis of primers: The sequence obtained from immature embryo expression profile sequencing (completed by Huada Genomics) of soybean variety Jilin 32 at three stages (20, 30 and 50 days) was input into NCBI for Blast comparison to obtain a Unknown cDNA sequence with high homology of ERF transcription factor protein sequence. According to the obtained unknown cDNA sequence, its nucleotide sequence is as described in Sequence NO.1, design synthetic primers, F: 5'-CTTCCTACTCCTCCCTTTCAC-3', R: 5'-CGTAGTAGTGTTCCCAGATGC-3'. Using the above cDNA as a template, carry out PCR amplification according to the follo...
Embodiment 2
[0028] Embodiment 2: the expression of GmERF6 gene in prokaryotic
[0029] Synthetic primers were designed according to the GmERF6 gene sequence, and Sac I and Hind III restriction sites were introduced into the upstream and downstream primers respectively, F: 5′-GAGCTCATGGCCCCAAGAGACAGC-3′; R: 5′-TTCGAATCAGGCAACCTCCGGGAG-3′. The pMD18-T-GmERF6 plasmid was used as a template for PCR amplification. After the amplified product was purified by an agarose gel DNA purification and recovery kit (purchased from TIANGEN), it was double-digested with Sac I and Hind III (all kinds of restriction enzymes were purchased from TaKaRa), and after recovery and purification, it was mixed with Similarly, the pET28a (purchased from Novagen) vector digested with Sac I and Hind III was used for ligation. The ligation product was transformed into Escherichia coli (E.coli) DH5α (purchased from Biovector), and after verification, it was transferred into the host strain Rosetta (DE3) (purchased from ...
Embodiment 3
[0033] Example 3: Expression pattern of GmERF6 gene under stress induction
[0034] Stress treatment of soybean Jilin 32: using Hoagland medium (Ca(NO 3) 2 4H 2 O 0.62g / L, KNO 3 0.34g / L, KH 2 PO 4 0.06g / L, NH 4 NO 3 0.053g / L, MgSO 4 ·7H 2 O 0.493g / L, MgCl 2 0.67mg / L, H 3 BO 3 0.38mg / L, ZnSO 4 ·7H 2 O 0.29mg / L, CuSO 4 ·5H 2 O 0.015625mg / L, FeSO 4 ·7H 2 O0.02785g / L, EDTA-Na 2 0.0373g / L, PH 5.7-5.8), 28°C, 16h light / 8h dark conditions to cultivate soybean hydroponic seedlings, take the four-leaf stage hydroponic seedlings and carry out the following treatments:
[0035] High-salt (NaCl) treatment: Place soybean seedlings in Hoagland culture solution containing 200mM NaCl, take samples after treatment for 0h, 1h, 2h, 5h, 10h and 24h, place them in liquid nitrogen quickly, and store them at -80°C for later use;
[0036] Drought treatment: Soybean seedlings were placed in Hoagland culture solution containing 20% PEG8000, samples were taken after treatment for 0...
PUM
Property | Measurement | Unit |
---|---|---|
molecular weight | aaaaa | aaaaa |
Abstract
Description
Claims
Application Information
- R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
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