Drought resistant relevant protein IbBT4 and coding gene and application thereof

A technology encoding genes and proteins, applied in drought-resistance-related protein IbBT4 and its encoding genes and applications, can solve problems such as extinction, solute leakage, and threats to agricultural safety

Active Publication Date: 2020-05-15
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

Method used

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  • Drought resistant relevant protein IbBT4 and coding gene and application thereof
  • Drought resistant relevant protein IbBT4 and coding gene and application thereof
  • Drought resistant relevant protein IbBT4 and coding gene and application thereof

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 test-tube plantlets 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 obtain...

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 resistant relevant protein IbBT4 and a coding gene and application thereof. The invention discloses the protein firstly. The protein is a protein of which the amino acid sequence is as shown in SEQID NO.1, or a fusion protein obtained through connection of an N end or/and C end of an amino acid sequence as shown in SEQID NO.1 with a protein tag. The invention further discloses the coding gene and the application of the protein. The invention finds the IbBT4 protein and the coding gene thereof, the coding gene of the IbBT4 protein is guided into arabidopsis thaliana, a transgenic arabidopsis thaliana plant overexpressing IbBT4 is obtained, the transgenic plant is subjected to drought stress treatment, the drought resistance is strengthened, and the droughtresistant relevant protein IbBT4 has important effects in a drought resistant process of plants, has important application value in research of increasing drought resistance of the plants, and has wide application space and great market prospects in agriculture 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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IPC IPC(8): C07K14/415C12N15/29C12N15/82A01H5/00A01H6/20
CPCC07K14/415C12N15/8273
Inventor 刘庆昌翟红何绍贞赵宁周媛媛
Owner CHINA AGRI UNIV
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