Three cotton ABF/AREB/ABI5/DPBF type transcription factors and coding genes and application thereof

A technology of transcription factors and coding genes, applied in the fields of application, genetic engineering, plant genetic improvement, etc.

Inactive Publication Date: 2011-05-18
THE INST OF BIOTECHNOLOGY OF THE CHINESE ACAD OF AGRI SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are no reports of ABF/AREB/ABI5/D

Method used

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  • Three cotton ABF/AREB/ABI5/DPBF type transcription factors and coding genes and application thereof
  • Three cotton ABF/AREB/ABI5/DPBF type transcription factors and coding genes and application thereof
  • Three cotton ABF/AREB/ABI5/DPBF type transcription factors and coding genes and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022]Example 1 Materials and methods for cloning genes encoding three cotton ABF / AREB / ABI5 / DPBF transcription factors GhABF2, GhABF3, and GhABI5

[0023] 1) Cotton material: The cotton material is selected from the optional variety Y18R.

[0024] 2) Strains: Escherichia coli E.coli DH5α, Agrobacterium LBA4404 and Pichia pastoris GS115.

[0025] 3) Vectors: pMD18-T, pG4AB, pBI121, pJawohl8-RNAi and pPIC9.0K.

[0026] 4) Tool enzymes and modification enzymes: various restriction enzymes and modification enzymes were purchased from TaKaRa Company, NEB Company and Fermentas Company.

[0027] 5) Chemical reagents: All chemicals are analytically pure at home and abroad.

[0028] 6) Primer synthesis: Synthesized by Shanghai Sangon Bioengineering Technology Service Co., Ltd.

[0029] 7) Sequencing: completed by BGI.

[0030] Experimental procedure

[0031] 1) Preparation of total RNA: The total RNA from the leaves of the land variety Y18R was extracted by the improved hot boric ...

Embodiment 2

[0069] Example 2 Analysis of Gene Structure Characteristics of GhABF2, GhABF3, and GhABI5 Transcription Factors

[0070] According to the obtained full-length cDNA sequences of GHABF2, GHABF3, and GHABI5 genes, primers were designed in combination with the translation initiation sites and termination sites predicted by bioinformatics, and PCR amplification was performed using cotton genomic DNA and cDNA as templates, respectively.

[0071] Primers used for PCR: GhABF2fp3: 5′-ATGGGGACTAACATGAACTT-3′

[0072] GhABF2rv3: 5′-CCAAGGACCGGTCTGGGTTC-3′

[0073] GhABF3fp3: 5′-ATGGGGTCTAATCTGAATTT-3′

[0074] GhABF3rv3: 5′-CCAAGGGCCTGTCAGTGTTC-3′

[0075] GhABI5fp3: 5′-ATGGTGGTTGAGAACTCTGA-3′

[0076] GhABI5rv3: 5′-TAATGGACCACTTAGATTTC-3′

[0077] The amplified fragments were recovered, connected to pMD18-T easy Vector, transformed into E.coli DH5α, and sent to Huada Genomics for sequencing after enzyme digesti...

Embodiment 3

[0078] Example 3 Analysis of the expression characteristics of GhABF2, GhABF3, GhABI5 transcription factor genes in cotton

[0079] 1) Template preparation: Total RNA was extracted from different cotton organ materials: roots, stems, leaves, calyxes, petals, stamens, ovaries, fibers, flower buds, and cotton bolls using the improved thermal boric acid method. The quality of the extracted RNA was determined by OD 260 / OD 280 Ratio and 0.7% agarose gel electrophoresis identification. Use it as a template to carry out reverse transcription reaction according to the following scheme: add 2 μg total RNA and 1 μL Oligo(dT) to a small PCR tube, incubate at 65°C for 15 minutes, then add 5×MMLV (RNase H - ) Buffer 5μL, dNTP (25mmol / L) 5μL, Ribonuclease Inhibitor 20U, MMLV 200U, and finally supplemented with DEPC-treated water to a total volume of 25μL, incubated at 42°C for 1h, inactivated MMLV in a water bath at 95°C for 5min, and obtained cDNA-20°C Save for later.

[0080] 2) Relat...

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Abstract

The invention discloses three novel cotton ABF/AREB/ABI5/DPBF type transcription factors GhABF2, GhABF3 and GhABI5, and coding genes and application thereof. Amino acid sequences of the GhABF2, GhABF3 and GhABI5 are shown as SEQ ID NO: 2, 5 and 8 respectively. Nucleotide sequences of the coding genes GhABF2, GhABF3 and GhABI5 are shown as SEQ ID NO: 1, 4 and 7 respectively. The invention also discloses the structural characteristics on genomic deoxyribonucleic acid (DNA) and a ribonucleic acid (RNA) editing law of reading frame regions of the coding genes GhABF2, GhABF3 and GhABI5. The invention also discloses the cotton expression characteristics and yeast expression characteristics of the three genes and the characteristic that the three genes can improve the drought resistance of arabidopsis thaliana. The genes provide gene resources for culturing a plant variety resistant to abiotic stress, and have great significance for improving the abiotic stress resistance of plants.

Description

technical field [0001] The invention relates to three ABF / AREB / ABI5 / DPBF type transcription factors derived from cotton, respectively named GhABF2, GhABF3, GhABI5, and their coding gene sequences. The invention also relates to the expression profiles of the three genes, the expression vector containing the genes, and the method for cultivating stress-resistant plants using the genes. Background technique [0002] Abiotic stresses such as drought and high salinity are important limiting factors for cotton yield and quality. In recent years, the intensification of changes in natural conditions has seriously affected the stability of cotton production, and to a certain extent affected the steady development of my country's cotton industry. Therefore, it is of great significance to carry out research on cotton stress resistance and improve its tolerance to abiotic stresses such as drought and high salinity through plant genetic engineering. [0003] ABA is the plant hormone wi...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12Q1/68C12N15/63C12N15/81C12N15/82C12R1/84
Inventor 张锐郭三堆梁成真
Owner THE INST OF BIOTECHNOLOGY OF THE CHINESE ACAD OF AGRI SCI
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