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Tobacco AKT1 gene and application

A gene and tobacco technology, applied in the tobacco AKT1 gene and application field, can solve the problems of less research on potassium ion channels and large potassium consumption.

Active Publication Date: 2019-02-22
GUIZHOU TOBACCO SCI RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Tobacco is a crop that consumes a lot of potassium. The potassium content of tobacco leaves is an important indi

Method used

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  • Tobacco AKT1 gene and application
  • Tobacco AKT1 gene and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Cloning of Example 1 Tobacco AKT1 Gene

[0032] Get 0.5g of tobacco (tobacco variety is K326) fresh leaves, use the Trizol method to extract the total RNA of tobacco cells, then use the cDNA synthesis kit of TaKaRa company to synthesize cDNA, and further use Primer5.0 software to design and obtain primers through artificial optimization. The primers include a forward primer and a reverse primer, and the nucleotide sequence of the forward primer is: 5'-ATGGGCAAAGAAAAATGGGC-3'; the nucleotide sequence of the reverse primer is 5'-TTAATTTTCTGAAGTACCAT-3', Use the synthesized cDNA as a template for PCR amplification. The PCR amplification system is 20 μL, including 10 μL of Premix ExTaq, 0.5 μL of 10 μM forward primer, 0.5 μL of 10 μM reverse primer, 1 μL of tobacco cell cDNA, ddH 2 O 8 μL; the reaction program of the PCR amplification is: pre-denaturation at 95°C for 5 minutes; denaturation at 95°C for 30 seconds; annealing at 55°C for 30 seconds; extension at 72°C for 2 mi...

Embodiment 2

[0034] Example 2 Biological Functional Analysis of Tobacco AKT1 Gene

[0035] 1. Purpose of the experiment

[0036] The biological function of tobacco AKT1 gene was verified by yeast functional complementation experiment.

[0037] 2. Experimental method

[0038] The potassium uptake-deficient yeast mutant strain R5421 was used as the recipient strain. Strain R5421 can be found in Maathuis F J Mand Sanders D 1996 Mechanisms of potassium absorption by higher plantroots. Physiol. Plant. 96, 158–168.

[0039] The T-vector connected with the tobacco AKT1 gene of Example 1 and the expression vector P416 (yeast free type shuttle expression vector, TEF constitutive promoter, CYC1 terminator, CEN6ARSH4 origin of replication, the screening marker in yeast is URA3, screening in Escherichia coli Marked as Amp. Carrier P416 can refer to Functional Expression of aω-3Fatty Acid Desaturase Gene from Glycine max in Saccharomyces cerevisiae) to carry out double enzyme digestion respectively ...

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Abstract

The invention relates to tobacco AKT1 gene and application. The sequences of the tobacco AKT1 gene and its coding protein are shown in SEQ ID NO:1 and 2 respectively. The AKT1 gene is cloned from tobacco for the first time, its biological function is verified by a functional complementation experiment of yeast. The AKT1 gene of tobacco has the function of promoting potassium ion absorption and transport.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to tobacco AKT1 gene and its application. Background technique [0002] Potassium ion channels are ion channels that allow potassium ions to specifically permeate the plasma membrane, while hindering the permeation of other ions, especially sodium ions. These channels are generally composed of two parts: one part is the channel region, which selects and allows potassium ions to pass through, while blocking sodium ions; the other part is a gating switch, which switches the channel on and off according to the signal in the environment. [0003] In the prior art, studies on potassium ion channel genes are extensive in the model plant Arabidopsis thaliana. For example, studies have shown that the Arabidopsis potassium channel gene AKT1 encodes an inward rectifier channel that can form homologous multimers, mainly in Arabidopsis thaliana Expressed in root epidermis and corte...

Claims

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Application Information

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IPC IPC(8): C12N15/29C07K14/415C12N15/82A01H5/00A01H6/82A01H6/20
CPCC07K14/415C12N15/8243
Inventor 任学良王仁刚李立芹鲁黎明王自力张洁
Owner GUIZHOU TOBACCO SCI RES INST
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