Soybean protein gmairp1 and its coding gene and application
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A protein and encoding technology, applied in the field of soybean protein GmAIRP1 and its encoding gene and application, can solve problems such as unsatisfactory, yield and quality decline, and restriction of soybean export
Active Publication Date: 2019-03-12
HARBIN NORMAL UNIVERSITY
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[0003] Soybean is an important economic crop and food crop in my country. Abiotic stresses such as drought and salinity have greatly reduced its yield and quality, which can neither meet the needs of the domestic people's lives, but also greatly limit the export of soybeans.
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Embodiment 1
[0049] Cloning and expression pattern of embodiment 1, GmAIRP1 gene
[0050] 1. Cloning of GmAIRP1 gene
[0051] Primers were designed using primer premier5.0, and two restriction endonuclease recognition sites, BamHI and SacI, were added to the 5′ ends of the primers respectively. The primer sequences were as follows:
[0056] The total RNA of Hefeng 45 soybean was extracted, cDNA obtained by reverse transcription was used as a template, and PCR amplification was carried out with GmAIRP1-P1 and GmAIRP1-P2 to obtain PCR amplification products.
[0057] The electrophoresis results of the PCR amplification products were as follows: figure 1 As shown, M: DL2000 molecular weight standard; 1 ~ 8: PCR amplification product, the target band with a size of about 640bp was obtained.
[0058] The above PCR amplification product was connected w...
Embodiment 2
[0070] Example 2, Functional Identification of GmAIRP1 Gene
[0071] 1. Construction of recombinant vector
[0072] The 642bp PCR product obtained in Example 1 was digested with BamHI and SacI, and connected with the expression vector PBI121 that had undergone the same digestion to obtain a recombinant vector.
[0073] After sequencing, the recombinant vector is a vector obtained by replacing the DNA fragment between the BamHI and SacI restriction sites of the expression vector PBI121 with the GmAIRP1 gene shown in Sequence 1 in the sequence listing, named pBI121-GmAIRP1.
[0074] 2. Preparation of recombinant bacteria
[0075] The purpose plasmid pBI121-GmAIRP1 was introduced into Agrobacterium EHA105 by three-parent hybridization, as follows:
[0076] (1) Inoculate the recipient Agrobacterium EHA105 on YEP solid medium containing 100mg / L rifampicin, culture at 28°C, and when a single colony grows, pick a single colony and inoculate it in the liquid medium, shake at 28°C t...
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Abstract
The invention discloses soybean protein GmAIRP1 and a coding gene and application thereof. The protein is one of the proteins as follows: the first protein is shown in a sequence 2 in a sequence table, and the second protein is formed in the mode that an amino acid sequence shown in the sequence 2 in the sequence table is subjected to substitution and / or deletion and / or addition of one or several amino acid residues, has the same function and is derived from the sequence 2. Experiments prove that a new gene GmAIRP1 is cloned, GmAIRP1-transferred tobacco is obtained by transferring the new gene GmAIRP1 into tobacco, the GmAIRP1-transferred tobacco and wild type tobacco are processed through a high-salt stress condition, and the growth vigor of the GmAIRP1-transferred tobacco is superior to that of the wild type tobacco; results of determination on the content of POD, CAT, MDA and praline of the two groups of plants show that the free radical scavenging capacity and the osmosis pressure adjusting capacity of the transgenic plant are totally higher than those of the wild plant, it is revealed that the GmAIRP1 is likely to participate in the anti-salt adjusting and controlling process of plants, and a foundation is laid for cultivating anti-salt plants.
Description
technical field [0001] The invention relates to the field of biotechnology, in particular to soybean protein GmAIRP1 and its encoding gene and application. Background technique [0002] Abiotic stresses such as salinity and drought are the main environmental factors that limit plant growth and development. These environmental factors lead to a series of physiological metabolic reactions in plants, manifested as reversible inhibition of metabolism and growth, and even cause irreversible damage in severe cases, leading to the death of the entire plant. During the long-term evolution, plants have gradually formed a series of defense mechanisms in response to adversity stress, among which the ubiquitin / 26S proteasome pathway is an important way to respond to stress. Ubiquitin ligase E3 determines the specific recognition of target proteins and plays a crucial role in the ubiquitination process. [0003] Soybean is an important economic crop and food crop in my country. Abiotic...
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