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Malus xiaojinensis Cheng et Jiang MxHA5 protein, and coding gene and application thereof

A gene and protein technology, applied in the field of Xiaojinhaitang MxHA5 protein and its coding gene and application, to achieve the effect of increasing iron deficiency resistance

Inactive Publication Date: 2012-07-11
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although many studies have demonstrated that H + The -ATPase gene has a great effect on improving the tolerance of plants, but it has not been seen about the plasma membrane H in the small golden crabapple. + -Report of ATPase gene

Method used

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  • Malus xiaojinensis Cheng et Jiang MxHA5 protein, and coding gene and application thereof
  • Malus xiaojinensis Cheng et Jiang MxHA5 protein, and coding gene and application thereof
  • Malus xiaojinensis Cheng et Jiang MxHA5 protein, and coding gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Embodiment 1, the acquisition of MxHA5 protein and its coding gene

[0054] 1. Obtaining the full length of the MxHA5 gene

[0055] Total RNA was extracted from leaves of Begonia chinensis and reverse transcribed into cDNA. Using cDNA as a template, using a primer pair composed of F1 and R1, PCR amplification was performed using a high-fidelity HiFi mix to obtain PCR amplification products.

[0056] F1: 5'-ATGGCCGGCGATAAGG-3';

[0057] R1: 5'-AACTGTATAATGTTGTTGAATTGTATCG-3'.

[0058] PCR amplification system (20μl): ddH 2 O 7.0 μl, 2×HiF Mix (0.5 U / μl) 10 μl, F1 (10 mM) 1.0 μl, R1 (10 mM) 1.0 μl and template 1.0 μl.

[0059] PCR amplification program: 94°C pre-denaturation for 5 minutes; then denaturation at 94°C for 30 s, annealing at 50°C for 30 s, and extension at 72°C for 3 min, a total of 33 cycles; finally, extension at 72°C for 10 min.

[0060] Perform 1% agarose gel electrophoresis on the PCR amplification product, see figure 1 A (Swimming lane M is a DNA ...

Embodiment 2

[0099] Example 2, the acquisition of transgenic yeast and the analysis of its resistance to heavy metal toxicity

[0100] 1. Obtaining of transgenic yeast

[0101] 1. Digest the recombinant T vector pEASY-MxHA5 constructed in Example 1 with restriction enzymes KpnI and XbaI, and recover the digested product (about 2965 bp).

[0102] 2. Digest pYES2.0 plasmid (yeast expression vector, Beijing Dingguo Changsheng Biotechnology Co., Ltd.) with restriction enzymes KpnI and XbaI, and recover the vector backbone (about 5800bp).

[0103] 3. Ligate the digested product of step 1 with the vector backbone of step 2 to obtain the recombinant expression vector pYES2.0-MxHA5.

[0104] 4. The recombinant expression vector pYES2.0-MxHA5 was introduced into the yeast strain BJ2168 using the yeast transformation kit (Beijing Pangeno Technology Co., Ltd.), and the transgenic yeast BJ2168 / pYES2.0-MxHA5 was obtained.

[0105] 5. Construction of control plasmid

[0106] (1) Digest pEASY-T1 sampl...

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Abstract

The invention discloses a malus xiaojinensis Cheng et Jiang MxHA5 protein, and a coding gene and application thereof. The protein provided by the invention is protein (a) or protein (b), wherein the protein (a) has amino acid sequences shown as a sequence 1 in a sequence table; and the protein (b) has an amino acid residue sequence which is derived from the sequence 1 by substituting and / or losing and / or adding one or more amino acid residues and is related with the P type H<+>-ATPase enzymatic activity in plant. The MxHA5 protein provided by the invention has an important effect of adjusting ion-deficient stressed plant, and the invention further provides good theoretical basis for the research on the ion-deficiency resistance of malus xiaojinensis Cheng et Jiang, and has significance to cultivation of stress-tolerant plant.

Description

technical field [0001] The invention relates to a small golden crabapple MxHA5 protein, its coding gene and application. Background technique [0002] Iron is an essential mineral element for the human body. Because plants are the main source of dietary iron for the world's population, a low iron content in plants will directly affect human health. According to the World Health Organization survey, nearly 2 billion people suffer from anemia. Premature births, congenital defects, mental retardation and decreased health indices are mainly caused by anemia. [0003] Studies have shown that plants can only absorb soluble iron-Fe(II) in the soil, but in alkaline soil (pH7.4-8.5), the concentration of soluble iron is less than 10 -10 mol·L-1, causing many plants to exhibit symptoms of iron deficiency stress. Mild iron deficiency in plants will lead to reduced chlorophyll synthesis and photosynthetic rate, severe iron deficiency will stop chlorophyll synthesis, new leaves will ...

Claims

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Application Information

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IPC IPC(8): C12N9/14C12N15/55C12N15/63C12N5/10C12N1/15C12N1/19C12N1/21A01H5/00
Inventor 韩振海槐心体王忆张倩张新忠许雪峰
Owner CHINA AGRI UNIV
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