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

A potassium transporter tpk1-1 from tobacco and its coding gene and application

A gene, tobacco technology, applied to potassium transporter TPK1-1 and its encoding gene and application field, can solve the problems of unknown tobacco TPK function and large potassium consumption, and achieve the effect of promoting potassium ion absorption and transport

Active Publication Date: 2021-08-27
GUIZHOU TOBACCO SCI RES INST
View PDF3 Cites 0 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
  • A potassium transporter tpk1-1 from tobacco and its coding gene and application
  • A potassium transporter tpk1-1 from tobacco and its coding gene and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1 Acquisition of TPK1-1 gene

[0046] Take 0.5 g of fresh tobacco leaves, extract the total RNA of tobacco cells by Trizol method, then use the cDNA synthesis kit of TaKaRa company to synthesize cDNA, and further use Primer 5.0 software to design and manually optimize primers, and the primers include forward primers and a reverse primer, the nucleotide sequence of the forward primer is: 5'-ATGGAGAAAGAGCCTCTTCT-3'; the nucleotide sequence of the reverse primer is 5'-TCAATGGTGGTGGCTTTCCA-3', using the synthesized 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; 35 cycles of denaturation at 95°C for 30s, annealing at 55°C for 30s, and extension at 72°C for 2 min.

[0049] After the PCR amplification ...

Embodiment 2

[0050] Example 2 The role of TPK1-1 gene in promoting potassium ion absorption and transport

[0051] The T-vector connected with the TPK1-1 gene described in Example 1 and the expression vector P416 were respectively subjected to double restriction digestion (the restriction sites are: SmaI and XhoI), the target gene and the expression vector P416 were recovered, and then connected with Enzyme ligation, transfer the ligated recombinant yeast expression vector into the competent cells of E. coli DH5α, carry out PCR amplification and digestion of the transformed E. coli single colony to verify whether the construction is successful, and the successful construction of the recombinant yeast expression vector will be carried out. Transfer to R5421.

[0052] The specific steps are as follows: streak the preserved R5421 yeast on the solid medium YPDA with an inoculating loop, and cultivate at 28°C for 12 hours; pick a single colony of R5421 yeast into an Ep tube, add 1 mL of YPDA me...

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 specifically discloses a potassium transporter TPK1-1 from tobacco, its coding gene and its application. The present invention provides the TPK1-1 gene isolated from tobacco for the first time. The full length of the TPK1-1 gene is 1287 bp. After functional verification, the TPK1-1 gene provided by the present invention is transferred into the potassium absorption-deficient yeast mutant strain R5421, and expresses the TPK1-1 gene. The recombinant yeast with the TPK1‑1 gene has regained the functions of potassium ion uptake and transport. Therefore, the TPK1-1 gene provided by the present invention has the function of promoting the absorption and transport of potassium ions.

Description

technical field [0001] The present invention relates to the field of genetic engineering, in particular to a potassium transporter TPK1-1 from tobacco and its encoding 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 generally consist of two parts: one is the channel region, which selects and allows potassium ions to pass through while blocking sodium ions; the other is the gated switch, which switches the channel on and off according to signals in the environment. [0003] The research on potassium channel genes in the prior art is relatively extensive in the model plant Arabidopsis thaliana. For example, studies have shown that the TPK / KCO channel in the model plant Arabidopsis thaliana includes a two-hole potassium channel TPK (also known as a tandem pore channel) and a Kir ...

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
Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/82A01H5/00A01H6/82A01H6/20
CPCC07K14/415C12N15/8243
Inventor 王仁刚任学良李立芹鲁黎明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