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Drought resistance related protein ibbt4 and its coding gene and application

A technology for encoding genes and proteins, applied in the field of drought resistance-related protein IbBT4 and its encoding genes and applications, can solve problems such as limiting growth and development, chloroplast damage, and reducing material accumulation

Active Publication Date: 2021-05-11
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Drought stress can cause plant cell membrane damage, solute leakage, and metabolic disorders; lead to stomatal closure, chloroplast damage, and reduced photosynthetic efficiency; affect enzymatic reactions, limit growth and development, and reduce material accumulation
Therefore, drought stress will cause crop yield reduction or even extinction, seriously threatening agricultural security.

Method used

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  • Drought resistance related protein ibbt4 and its coding gene and application
  • Drought resistance related protein ibbt4 and its coding gene and application
  • Drought resistance related protein ibbt4 and its coding gene and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0076] Embodiment 1, the acquisition of IbBT4 gene

[0077] 1. Acquisition of cDNA template

[0078] The total RNA of the sweet potato line Xushu 55-2 tube seedlings was extracted with a plant total RNA extraction kit, and the first-strand cDNA was reverse-transcribed from the total RNA with a PrimeScriptTM RT reagent Kit with gDNA Eraser kit.

[0079] 2. Using the cDNA obtained in step 1 as a template, design and artificially synthesize primers 3GSP-1 and 3GSP-2 according to the EST sequence, use the RACE method to amplify a 3′-RACE fragment of about 800bp, and clone the 3′-RACE fragment and The vector pMD19-T was ligated to obtain recombinant plasmid 2. The recombinant plasmid 2 was sequenced to obtain the nucleotide sequence of the 3'-RACE fragment. The primer sequences are as follows:

[0080] 3GSP-1: 5′-CGAAAACTTTAGGCAGCAGG-3′

[0081] 3GSP-2: 5'-AGTCCGTTTCCTTCACCGAA-3'

[0082] 3. Design and artificially synthesize primers 5GSP-1 and 5GSP-2, use the cDNA obtained in...

Embodiment 2

[0089] Example 2. Application of IbBT4 protein in improving plant drought resistance.

[0090] 1. Construction of recombinant plasmid pCAMBIA super1300-IbBT4-GFP

[0091] 1. Artificially synthesize the double-stranded DNA molecule shown in positions 1 to 1116 from the 5' end of SEQ ID NO.2. Using the double-stranded DNA molecule as a template and OE-F-Xba I:

[0092] 5′-GC TCTAGA ATGGGTAAGCTTTCGGATTC-3' (underlined is the recognition sequence of restriction endonuclease Xba I) and OE-R-Pst I: 5'-AA CTGCAG TGTTGCTTCAACTGAGAAAAAT-3' (the underline is the recognition sequence of the restriction endonuclease PstI) is used as a primer for PCR amplification to obtain a double-stranded DNA molecule containing the restriction endonuclease XbaI at the N-terminus and the restriction endonuclease PstI at the C-terminus .

[0093] 2. Digest the vector pCAMBIA super1300-GFP with restriction endonucleases Xba I and Pst I, and recover the vector backbone 1 of about 10783 bp.

[0094] ...

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Abstract

The invention discloses a drought-resistance-related protein IbBT4, its coding gene and application. The present invention firstly discloses a protein, which is a protein whose amino acid sequence is shown in SEQ ID NO.1 or a fusion protein obtained by linking protein tags at the N-terminal or / and C-terminal of the amino acid sequence shown in SEQ ID NO.1. The invention further discloses the coding gene of the above protein and its application. The present invention discovers the IbBT4 protein and its coding gene, introduces the coding gene of the IbBT4 protein into Arabidopsis thaliana, obtains transgenic Arabidopsis plants overexpressing IbBT4, and carries out drought stress treatment on the transgenic plants, and its drought resistance is enhanced, and the plant It plays an important role in the process of drought resistance, has important application value in the study of improving plant drought resistance, and has broad application space and market prospects in the agricultural field.

Description

technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to a drought-resistance-related protein IbBT4 and its coding gene and application. Background technique [0002] Crops are affected by a variety of biotic or abiotic stresses in actual production, such as salt and drought, light and temperature, heavy metals, pests and diseases, etc. The shortage of global water resources also makes agricultural production face increasingly severe challenges. China's arid land accounts for 50% of the country's land area, and China's per capita water resources are only 1 / 4 of the world's average level. Current status limited by water resources. With the rapid expansion of population and global warming, the shortage of water resources has become one of the main challenges facing agriculture. China's agricultural production is increasingly affected by drought disasters, and food security is facing severe challenges. For the sustainable develop...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/82A01H5/00A01H5/10A01H6/20
CPCC07K14/415C12N15/8273
Inventor 刘庆昌翟红何绍贞赵宁周媛媛
Owner CHINA AGRI UNIV