Gene sequence of SOD (Superoxide Dismutase) protein of Prunus pedunculatacv.Wulate and application of SOD protein
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A technology of peduncle almond and protein, applied in the field of Urad peduncle almond SOD gene and its encoded protein sequence
Active Publication Date: 2014-10-29
INNER MONGOLIA AUTONOMOUS REGION ACAD OF AGRI & ANIMAL HUSBANDRY SCI
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So far, the SOD gene sequence obtained from Urad peduncle almond has not been reported
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Embodiment 1
[0022] Embodiment 1, the acquisition of the SOD protein coding gene of Urad peduncle almond
[0023] 1.1 Extraction of RNA
[0024] RNA extraction was carried out with reference to the method of Zhang Jinsong et al. (Zhang Jinsong, et al. Chinese Science, Series B, 1995, 25 (5): 659-669), the root of Urad peduncle almond was ground in liquid nitrogen, suspended in the extract (4mol Sodium in / L guanidine thiocyanate, 5g / L lauryl sarcosine, 25mmol / L sodium citrate, 0.1mol / L β-mercaptoethanol), the mixture was extracted with acidic phenol and chloroform, and the supernatant was added without Precipitate the total RNA with water and ethanol, then suspend the RNA with 4mol / L LiCl for precipitation, extract with chloroform once, then precipitate the total RNA with sodium acetate / ethanol, and finally dissolve the RNA in water and store it at -20°C (for later use, use 0.8% Agarose assay for RNA integrity.
[0025] 1.23'RACE sequence acquisition:
[0026] Using the peach SOD gene s...
Embodiment 2
[0034] Example 2, Sequence and function analysis experiment of Urad peduncle almonds SOD protein
[0035] 2.2 Bioinformatics analysis:
[0036] Using BLAST and DNAMAN software to compare and analyze the SOD protein sequence of Urad peduncle almond, the results show that Urad peduncle SOD has great homology with the SOD of peach, rubber tree and wild strawberry (such as Figure 4 , Figure 5 shown).
[0037] 2.2 Transgenic analysis:
[0038] 2.2.1 Construction of expression vector
[0039] Design 1 pair of primers for the construction of plant expression vectors
[0042] (wherein the underlined parts represent XbaI, BamHI restriction sites respectively).
[0043] The construction process of the plant expression vector: extract the total RNA of almond leaf, and use oligo(dT) 18 for reverse transcription. The reverse transcription product was PCRed with...
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Abstract
The invention discloses a SOD (Superoxide Dismutase) protein of Prunus pedunculatacv.Wulate and a coding gene cloning method and a sequence thereof. According to the SOD protein and the coding gene cloning method and the sequence thereof disclosed by the invention, the full-length of the SOD gene of the Prunus pedunculatacv.Wulate is cloned by the RACE method and the coding protein of the SOD gene is analyzed to discuss the effect of the gene in improving oxidation resistance, salt stress and the like of transgenic plants. According to the SOD gene of the Prunus pedunculatacv.Wulate as a potential stress resistance candidate gene, cloning and functional analysis of the full gene of the SOD gene as well as genetic transformation to plants have certain meaning for culturing high stress resistance plant varieties. The SOD gene of the Prunus pedunculatacv.Wulate obtained by the method disclosed by the invention exerts an important effect in researching the antioxidation mechanism of the Prunus pedunculatacv.Wulate and the stress resistance of rosaceous plants.
Description
technical field [0001] The invention relates to a Urad peduncle almond SOD gene and its encoded protein sequence, gene cloning method and application. Background technique [0002] Superoxide dismutase (Superoxide dismutase, referred to as SOD) is an enzyme that can catalyze the conversion of superoxide into oxygen and hydrogen peroxide through the dismutation reaction, effectively prevent their damage to organisms, and almost participate in the It is a physiological and biochemical reaction to adversity and is a very important antioxidant enzyme in organisms. It widely exists in various animals, plants and microorganisms. [0003] According to the different metal ions combined by SOD, it can be divided into three types: Fe-SOD, Mn-SOD and Cu / Zn-SOD. At present, scholars at home and abroad have successively cloned the full-length cDNA sequence or fragment sequence of the gene from poplar, sweet potato, cassava, peach and other crops. [0004] Prunus pedunculatacv. Wulate ...
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