Peanut S-adenosylmethionine synthetase gene AhSAMS and protein and application

A technology of adenosylmethionine and synthetase, applied in the field of biological genes, can solve the problem of lack of regulation of peanut SAMS gene and the like, and achieve the effect of improving peanut yield

Inactive Publication Date: 2016-06-15
BIOTECH RES CENT SHANDONG ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In order to solve the problem that there is no report on the peanut SAMS gene and its regulatory role in plant stress physiology and aging physiological process in the above prior art, the application provides the peanut S-adenosylmethionine synthetase gene AhSAMS

Method used

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  • Peanut S-adenosylmethionine synthetase gene AhSAMS and protein and application
  • Peanut S-adenosylmethionine synthetase gene AhSAMS and protein and application
  • Peanut S-adenosylmethionine synthetase gene AhSAMS and protein and application

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Embodiment 1

[0033] The present invention obtains the peanut AhSAMS gene described in the present invention through the following experimental methods.

[0034] 1. Yeast two-hybrid

[0035] 1. Reagents used in the experiment

[0036] The kit used in the yeast two-hybrid experiment: BDMatchmakerTMLibraryConstruction & ScreeningKits was purchased from Clontech Company. The reagents used in this experiment refer to the product instructions.

[0037] 2. Construction of pGBKT7(BD) expression vector pGBKT7-CaM

[0038] After extraction of peanut leaf RNA, the cDNA was obtained by reverse transcription, and the 5' primer 1 (5'-CATATGATGGCGGATCCGCTCACC-3') containing the NdeI site and the 3' primer 2 (5'-CTGCAGCTAGGCCATCATGACCTTG -3′), amplify the full length of the calmodulin gene AhCaM in peanut leaves (Accessionnumber: AY517930) from the peanut cDNA, and verify the correctness of the amplified fragment by sequencing. The pGBKT7-CaM vector was constructed by digestion and ligation.

[0039]...

Embodiment 2

[0125] Example 2: Interaction verification of AhSAMS and AhCaM

[0126] 1. Verification of yeast two-hybrid system

[0127] According to the sequence obtained by screening and sequencing, primer 3 (5'-TCTAGAATGGCAGCAGAGACTTTCC-3') and primer 4 (5'-GGATCCTTAGGCCTTCTCCCACTT-3') were designed to obtain the full-length AhSAMS gene through PCR amplification. Construct pGBKT7-AhSAMS and pGADT7-AhCaM and empty vector as controls, and co-transform AhSAMS and AhCaM into yeast strain Y 2 For HGold, see steps 3 and 4 in Example 1 for the conversion method. Pipette 1 μL of bacterial liquid onto SD–Leu / –Trp, SD–Ade / –His / –Leu / –Trp / X / A solid screening media respectively, observe the growth and draw a graph.

[0128] Transform the recombinant plasmid into yeast cell Y 2 HGold, observe cell growth in corresponding selection media. Such as Figure 4 As shown, compared with the control strain, we found that pGADT7-AhCaM and pGBKT7(BD) empty vector strains could survive in SD / -Trp / -Leu mediu...

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Abstract

The invention provides a peanut S-adenosylmethionine synthetase gene AhSAMS and protein and application thereof, and the sequence of the gene is shown in the first sequence in a sequence table. Peanut S-adenosylmethionine synthetase which is interactive with calmodulin protein (CaM) is screened out by screening a protein library, and the genetic sequence and the protein sequence of the peanut S-adenosylmethionine synthetase are obtained. The functions of the peanut S-adenosylmethionine synthetase are determined through further experiments, the different expression conditions of the peanut S-adenosylmethionine synthetase in adversity stress are obtained through draught, high salt, ABA and high temperature highlight stress treatment, and a factual basis is supplied to following further utilization of the peanut S-adenosylmethionine synthetase gene, so that the regulation and control capacity of peanuts to adversity is conveniently improved by further utilizing Ca<2+> and the gene AhSAMS.

Description

technical field [0001] The invention relates to the technical field of biological genes, in particular to peanut S-adenosylmethionine synthetase gene AhSAMS, peanut S-adenosylmethionine synthetase protein, and peanut S-adenosylmethionine synthetase Application of gene AhSAMS and peanut S-adenosylmethionine synthetase protein. Background technique [0002] Peanut is an oilseed and economic crop widely grown in the world, and it is one of the most important oilseed crops in the world. China is the largest peanut producing and exporting country in the world. The peanut planting area ranks seventh among crops in my country, and the agricultural output value ranks fifth. The total annual output has reached more than 15 million tons, accounting for 43% of the world's total peanut production. %, the export volume accounts for more than 40% of the world. However, although my country's oil crop production has developed rapidly, it still cannot keep up with the continuous improvement...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/54C12N9/10A01H5/00
CPCC12N9/1085C12N15/8273C12N15/8293C12Y205/01006
Inventor 杨莎李新国万书波郭峰张佳蕾耿耘孟静静黄超
Owner BIOTECH RES CENT SHANDONG ACADEMY OF AGRI SCI
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