Potassium Channel Protein, Its Encoding Gene and Application

A technology of R. longarum and encoding gene, which is applied in the field of external rectifier potassium channel protein of R. longarum and its encoding gene and application field, and can solve the problem of unknown biological function of SKOR and the like.

Active Publication Date: 2022-01-11
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the research on SKOR of E. elongatum has not been reported in the prior art. Whether SKOR exists in E. elongatum and what biological functions of SKOR are unknown

Method used

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  • Potassium Channel Protein, Its Encoding Gene and Application
  • Potassium Channel Protein, Its Encoding Gene and Application
  • Potassium Channel Protein, Its Encoding Gene and Application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1 Cloning and sequence analysis of EeSKOR outer rectifier potassium channel protein gene EeSKOR of Echinopsis elongatum

[0047] Degenerate primers (P1, P2) were designed, and the cDNA synthesized by reverse transcription of the total RNA of the young roots of E. elongatum was used as a template to amplify the conserved core cDNA fragment of E. elongatum EeSKOR by PCR. After sequencing, the core fragment was found to be 555bp ( figure 1 a).

[0048] According to the 5'-RACE and 3'-RACE methods of the Clontech SMARTer RACE kit manual, 5'-cDNA and 3'-cDNA were synthesized respectively; based on the core sequence of the E. elongatum EeSKOR gene, DNAMAN 8.0 and Primer5.0 were used to The software designed the 5'-RACE outer primer P3, the Z-type primer P4 and the 3'-RACE outer primer P5, and the Z-type primer P6; the 3'-RACE amplification product was sequenced to 1033bp ( figure 1 b), the 5'-RACE amplification product is sequenced to be 1136bp ( figure 1 c).

[00...

Embodiment 2

[0062] Example 2 Effects of different concentrations of salt treatment on the expression level of EeSKOR

[0063] In order to ascertain the expression level and changes of EeSKOR gene in the roots, sheaths and leaves of E. elongatum under salt stress, qRT-PCR forward primer P7 (5'-TACGGAGGCTGCTCAGGTTT-3') and reverse primer P8 (5' -CGCATCTCCTCGCTTCATC-3'), the PCR product length is 189bp; internal reference gene Actin real-time fluorescent quantitative PCR forward primer P9 (5'-CTTGACTATGAACAAGAGCTGGAAA-3') and reverse primer P10 (5'-TGAAAGATGGCTGGAAAAGGA-3'), PCR product The length is 139bp, and the expression levels of EeSKOR gene in roots, leaf sheaths and leaves of 4-week-old Etypyrum elongatum were treated with different concentrations of NaCl (0, 25, 50, 100, 150 and 200mM) for 24h.

[0064] The results showed that with the increase of NaCl treatment concentration (25-100mM), the expression level of EeSKOR gene in roots showed an increasing trend, while the expression le...

Embodiment 3

[0065] Example 3 Construction of Etoxypyrum elongatum EeSKOR gene plant expression vector

[0066] According to the requirements of Clontech Infusion seamless connection technology, Nco was introduced at both ends of the primers upstream (P11: 5'-ACTCTTGACCATGGTATGGAGAGGGAGATTGTAGCAGAG-3') and downstream (P12: 5'-TTTACCCTCAGATCTCTACTGATCGGCTGCAACAGCAG-3') of the EeSKOR gene ORF frame. I and Bgl II restriction site, through RT-PCR amplification, by 1.2% gel electrophoresis detection target fragment PCR product ( Figure 5 ). Then, the restriction enzyme site on the pCAMBIA1301 vector was double-digested with Nco I and Bgl II, and the large fragment was recovered, named pCAMBIA1301-A; according to the corresponding procedure of Clontech Infusion seamless linking technology, the target gene EeSKOR was inserted into the linear The recombined plasmid pCAMBIA1301-35S-EeSKOR-Nos was obtained from the surface vector (pCAMBIA1301-A) of the plant, and then the specific band of about 29...

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Abstract

The present invention relates to the field of biotechnology, in particular to a rectifying potassium channel protein outside of Triticum aurantiae, and its encoding gene and application. The present invention clones the outer rectifier potassium channel protein EeSKOR of R. longa, and its amino acid sequence is shown in SEQ ID NO.1. In the present invention, it is found that EeSKOR can increase the biomass and plant height of plants in a high-salt environment, and over-expression of the EeSKOR gene under salt treatment significantly reduces the H content of transgenic tobacco plants. 2 O 2 and MDA content, increased SOD activity and chlorophyll content, significantly decreased Na in transgenic plants + concentration, increasing its K in the body + concentration. EeSKOR can be used to improve the growth performance of plants under high salt stress and improve the salt tolerance of plants, which is of great significance for the cultivation of new varieties of salt-tolerant plants.

Description

technical field [0001] The invention relates to the field of biological technology, in particular to the outer rectifier potassium channel protein of Echinopsis elongatum and its encoding gene and application. Background technique [0002] When plants are faced with abiotic stresses such as salt stress, the response involves self hypersensitivity that culminates in programmed cell death. K + / Na + If the ratio is too low, the plant degrades its contents by autophagy, or autophagy in aleurone cells, or degradative hydrolases released during the formation of tubular molecules in cell protoplasts. Therefore, high K in plants + / Na + Ratio, related to plant salt tolerance, and related to plant growth and development. K + channel and K + The transporter is the plant body for K + An important part of the absorption, transport and distribution of Gaymard et al. found that ABA treatment and acidic conditions could reduce the expression of SKOR. AtSKOR isolated from Arabidop...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/84A01H5/00A01H6/82
CPCC07K14/415C12N15/8273C12N15/8261
Inventor 郭强孟林周妍彤毛培春田小霞郑明利
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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