Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Potassium transport protein TPK1 from tobacco and coded gene and application thereof

A gene, tobacco technology, applied to potassium transporter TPK1 and its encoding gene and application field, can solve the problems of unknown tobacco TPK function, large potassium consumption, etc., and achieve the effect of promoting potassium ion absorption and transport

Active Publication Date: 2019-02-19
GUIZHOU TOBACCO SCI RES INST
View PDF5 Cites 2 Cited by
  • 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 indicator to measure the quality of tobacco leaves. At present, there are few studies on potassium ion channels in tobacco, and the function of tobacco TPK is unknown.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Potassium transport protein TPK1 from tobacco and coded gene and application thereof
  • Potassium transport protein TPK1 from tobacco and coded gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Obtaining of embodiment 1 TPK1 gene

[0046] Get 0.5g of fresh tobacco 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 the Primer5.0 software to design and obtain primers through artificial optimization. The primers include forward primers and a reverse primer, the nucleotide sequence of the forward primer is: 5'-ATGACTAACAGAGACAAAATCCAGC-3'; the nucleotide sequence of the reverse primer is 5'-TCACCTTCCTGTCCAAGACGATTGG-3', using the synthetic cDNA as a template , for PCR amplification.

[0047] The PCR amplification system is a 20 μL system, including: Premix ExTaq 10 μL, 10 μM forward primer 0.5 μL, 10 μM reverse primer 0.5 μL, tobacco cell cDNA 1 μL, ddH 2 08 μL.

[0048] The PCR amplification reaction program was: pre-denaturation at 95°C for 5 min; denaturation at 95°C for 30 s, annealing at 55°C for 30 s, extension at 72°C for 2 min, and 35 cycles.

[0...

Embodiment 2

[0050] Example 2 The role of TPK1 gene in promoting the absorption and transport of potassium ions

[0051] The T-vector connected with the TPK1 gene described in Example 1 and the expression vector P416 were subjected to double enzyme digestion (restriction sites: SmaI and XhoI), and the target gene and expression vector P416 were recovered, and then ligated with ligase , transfer the ligated recombinant yeast expression vector into Escherichia coli DH5α competent cells, carry out PCR amplification and enzyme digestion on the transformed Escherichia coli single colony to verify whether the construction is successful, and transfer the successfully constructed recombinant yeast expression vector into into R5421.

[0052] The specific steps are as follows: Streak the preserved R5421 yeast on the solid medium YPDA with an inoculation loop, culture at 28°C for 12 hours; pick a single colony of R5421 yeast in the Ep tube, add 1mL of YPDA culture solution and vortex; Transfer all t...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention relates to the field of genetic engineering, and particularly discloses a potassium transport protein TPK1 from tobacco and a coded gene and application thereof. A TPK1 gene separated from the tobacco is provided for the first time, the full length of the TPK1 gene is 1062 bp, functional verification proves that after the provided TPK1 gene is transferred to a potassium uptake defected type yeast mutant strain R5421, the function of potassium ion absorbing and transferring is regained by a recombinant yeast expressing the TPK1 gene, and therefore the provided TPK1 gene has the function of promoting potassium ion absorbing and transferring.

Description

technical field [0001] The invention relates to the field of genetic engineering, in particular to a potassium transporter TPK1 from tobacco and its coding gene and application. Background technique [0002] Potassium ion channels are ion channels that allow potassium ions to specifically permeate the plasma membrane, while blocking 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 TPK / KCO channel in the model plant Arabidopsis includes two-pore potassium channel TPK (also known as tandem pore channel) and a Kir -type channel. T...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C07K14/415C12N15/29C12N15/82A01H5/00A01H6/20A01H6/82
CPCC07K14/415C12N15/8243
Inventor 任学良王仁刚鲁黎明李立芹郭玉双张洁王自力林世锋
Owner GUIZHOU TOBACCO SCI RES INST
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com