Gene sequence of glutathionetransferase of vinca rosea
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A technology of glutathione and periwinkle, applied in the direction of transferase, genetic engineering, plant gene improvement, etc., can solve the problem of undiscovered periwinkle GST protein gene sequence and amino acid sequence, etc.
Inactive Publication Date: 2005-10-26
NORTHEAST FORESTRY UNIVERSITY +1
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[0010] After searching, no published or reported gene sequence and amino acid sequence of GST protein of Vinca
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Embodiment 1
[0021] Example 1 Cloning of the full-length gene of periwinkle drought stress-related gene glutathione transferase
[0022] Step 1, Plant Material Processing
[0023] The periwinkle seedlings were sown in perlite, cultivated in a light incubator, and watered every day. The vinca plants of 1 month old were naturally dehydrated in the light incubator, and the water potential and osmotic potential of the leaves were measured every hour. Up to -1.0MPa is the standard material. The leaves were quickly frozen in liquid nitrogen and immediately placed in a -70°C ultra-low temperature freezer.
[0024] Step 2, extraction of total RNA and purification of mRNA
[0025] 1. Use Trizol (Gibco) to extract total RNA in one step, according to the kit instructions. Total RNA was quantified and analyzed by denaturing electrophoresis.
[0026] 2. Purification of mRNA: Oligotex mRNA Purification Kit (Qiagen) was used to purify mRNA from total RNA and quantify it.
[0027] Step 3, λZAP Expre...
Embodiment 2
[0104] Example 2 Sequence information and homology analysis of periwinkle GST
[0105] The following part is the nucleotide sequence of the cloned Vinca GST:
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Abstract
A vinca rosea Glutathione S-transferase gene sequences, and it coding product amino acid sequence. It relates to a kind of new gene sequence, characterized in that: this gene is 929bp, at the point of 56bp there is the start coding triplet ATG, 718bp the ending coding triplet TGA, 910bp additional signal polyA. The gene sequence has not any inner gene, 663bp completely disclosed code-reading frame, code 220 amino acid. Nucleotide sequence and Capsicum Chinese gene GST1 has reached 67.7% homeotic source, and Euphorbia esula gene GST 57.7 homeotic source. This invention also provides a method to gain a full length of forced gene. First we enrich the anti-dry expressing gene in a moderate dry condition; then we establish full length genomic library of cDNA and detecting order, obtain the anti-dry related full length forcing gene in most short period. This anti-dry gene straightly links to eukaryotes expressing carrier pCMV plasmid, detected of the order by t3, t7 primer, filtered by eukaryotes expressing. This invention distills the full length gene of glucoside peptide transferase. This gene has a good anti-oxidate function, increases express in adverse circumstance forces in the process, the gene eliminates the free radical of oxidation. This gene colon expands the gene source of plant anti-dry, and provides a more rich and good usable gene for enhancing the anti-dry function and meliorating breed.
Description
[0001] Field [0002] The invention belongs to the field of biological genes, and in particular relates to the gene sequence of periwinkle drought stress-related gene glutathione transferase. Background technique [0003] Due to the imbalance of ecological balance, more than one-third of the earth's land area belongs to arid and semi-arid areas, so it is urgent to develop and utilize arid lands. It is an economical and effective way to use arid land to study the drought-resistant mechanism of plants, apply molecular biology methods to clone drought-resistant genes, and use genetic engineering techniques to breed drought-tolerant species. [0004] Under adversity conditions such as salt and drought, the decline of active oxygen scavenging or antioxidant capacity in plants is the main reason for the large accumulation and increase of active oxygen. The accumulation of reactive oxygen species can initiate the peroxidation of unsaturated fatty acids in membrane lipids, eventually...
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