Thinopyrumponticum tonoplast Na+/H+ reverse transport protein 2 gene and use thereof

A technology of antiporter and tonoplast membrane, applied in the field of Na+/H+ antiporter 2 gene

Inactive Publication Date: 2005-11-23
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no report about the gene of Na+ / H+ antiporter 2 on the tonoplast membrane of Agropyresis and its application in plant transgenic engineering breeding

Method used

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  • Thinopyrumponticum tonoplast Na+/H+ reverse transport protein 2 gene and use thereof
  • Thinopyrumponticum tonoplast Na+/H+ reverse transport protein 2 gene and use thereof
  • Thinopyrumponticum tonoplast Na+/H+ reverse transport protein 2 gene and use thereof

Examples

Experimental program
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Effect test

Embodiment Construction

[0067] 1. High wheatgrass Na + / H + Cloning of Partial Fragment of Antiporter 2 Gene

[0068] Trizol method to extract total RNA from wheatgrass seedlings germinated for about 2 weeks, PowerScript TM The first-strand cDNA was synthesized by reverse transcription with reverse transcriptase, and PCR was carried out with degenerate primers: P1: 5`-GAT TCT GTW TGC ACM YTR CAG GT-`3, P2: 5`-CAT KAG MCC AGC CCA CCA WAT-`3 Amplify, obtain an amplified fragment, clone and sequence.

[0069] Amplify the target gene fragment according to the following conditions:

[0070] Reaction system: 2×GC buffer I 25μl, 10mM dNTP 3μl, 10μM primer 1 and primer 2 1.5μl each, DNA template 250ng, LA GC Taq enzyme 1μl, add water to make up to 50μl.

[0071] The reaction program was: pre-denaturation at 94°C for 3 min, followed by 36 cycles, each cycle including denaturation at 94°C for 40 s, renaturation at 59°C for 1 min, and extension at 72°C for 1 min. Finally, it was kept at 72°C for 10 min.

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Abstract

The invention discloses a thinopyrumponticum tonoplast Na+ / H+ reverse transport protein 2 gene, and the application of the gene sequence in improving crop and forage grass' salt resistant character by means of biological technique. The provided gene is a member of the Na+ / H+ antiporter family on the Thinopyrumponticum vacuole membrane, the gene shows positive relativity to the salt tolerance of the Thinopyrumponticum. The disclosed gene sequence can be employed to clone the gene and construct pronucleus expression carrier. The mass expression in procaryotic cells can produce Na+ / H+ antiporter 2 for protein structural research, functional research and antigen preparation. After cloning the gene based on the gene sequence, various pronucleus expression carriers can be constructed, by leading the gene into crops and forage grass, strains / lines with high salt tolerance property can be cultivated.

Description

technical field [0001] The present invention relates to the tonoplast membrane Na + / H + The antiporter 2 gene (TeNHX2) belongs to the field of biotechnology. technical background [0002] Strategies for plants to eliminate Na+ toxicity include: reducing Na+ uptake, Na+ efflux and Na+ compartmentalization. Na+ uptake is a complex process. Na+ efflux and compartmentalization are indirect active transport processes, mainly regulated by Na+ / H+ antiporters. Na + / H + Antiporter (Na + / H + Antiporter or exchanger, NHA or NHE) is a transport protein ubiquitous in the membrane system of bacteria, yeast, algae, animals and higher plants, involved in the pH, Na + Life activities such as concentration regulation and cell volume change. Since Blumwald and Plooe first discovered the Na+ / H+ antitransport activity on the tonoplast membrane of sugar beet root storage tissues in 1985, people have successively discovered the Na+ / H+ reverse transport activity in halophytes such as A...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29
Inventor 夏光敏刘恒薛哲勇
Owner SHANDONG UNIV
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