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Transcription factor of DREB class in cotton, its coding gene, and application

A transcription factor, cotton technology, applied in the fields of application, genetic engineering, plant gene improvement, etc., can solve the problems such as the analysis of the functions of members of the A-6 group

Inactive Publication Date: 2006-12-06
TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, so far, except for ZmDBF1, the functions of other A-6 group members have not been resolved, and there is no experimental evidence that there are genes similar to ZmDBF1 in other plants.

Method used

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  • Transcription factor of DREB class in cotton, its coding gene, and application
  • Transcription factor of DREB class in cotton, its coding gene, and application
  • Transcription factor of DREB class in cotton, its coding gene, and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Embodiment 1, the cloning of cotton DREB class transcription factor coding gene GhDBP2

[0045] The cDNA sequence of the cotton DREB transcription factor coding gene GhDBP2 was cloned by combining bioinformatics and conventional PCR. and DBF2 are involved in rab17regulation through the drought-responsive element in an ABA-dependent pathway. Plant J. 2002, 30: 679-689) encoded amino acid sequence was searched cotton EST database (http: / / www.tigr. org / ), the result was a sequence (TC18189), which was spliced ​​from several EST sequences from Upland cotton and Sea island cotton, with a translation start codon and a stop codon, indicating that the sequence contained the full-length open Reading frame (ORF), sequence alignment results show that the amino acid residue sequence encoded by this sequence has a high similarity with the amino acid residue sequence of ZmDBF1. In order to verify that the cDNA sequence also exists in upland cotton (Gossypium hirsutum), a pair of pri...

Embodiment 2

[0046] Example 2, Sequence Analysis of GhDBP2 Protein and Homology Evolution Analysis of DNA Binding Domain

[0047] 1. Sequence analysis of GhDBP2 protein

[0048] Sequence analysis of the amino acid residue sequence of GhDBP2 revealed that the GhDBP2 protein contains an AP2 / ERF domain that is conserved among DREB subfamily transcription factors, namely amino acid residues 160-217 from the amino terminal (N terminal) (The 587th-760th base from the 5' end in SEQ ID №: 2 is its coding sequence), a total of 58 amino acids; there is an acidic region at its C-terminus, that is, from the 278th-350th amino-terminal Amino acid residues (bases 941-1159 from the 5' end in SEQ ID №2 are its coding sequence), the acidic region may function as a transcriptional activation region; behind the DNA binding region is the amphipathic nuclear localization Signal sequence (NLS), that is, amino acid residues at position 257-273 from the amino terminal (in SEQ ID No.: 2, bases at position 878-928 ...

Embodiment 3

[0053] Example 3, Analysis of the binding characteristics of GhDBP2 and DRE elements

[0054] 1. Preparation of GhDBP2 AP2 / ERF domain fusion protein with His-tag (6×His)

[0055] 1. Construction of GhDBP2 AP2 / ERF domain expression vector

[0056] Using GhDBP2 cDNA as a template, primer P1 (upstream primer): 5'-CG GGATCC ATGGTTAGCTTTCTTTGC-3' (the underlined base is the restriction endonuclease BamH I recognition site) and P2 (downstream primer): 5'-CG GAATTC Under the guidance of TCAAGCGTCAACAGAGGA-3' (the underlined base is the restriction endonuclease EcoRI recognition site), PCR amplified the GhDBP2 AP2 / ERF domain coding sequence. After the reaction, the PCR amplified product was detected by 1.2% agarose gel electrophoresis, and the target fragment of about 300bp was recovered and purified. After it was digested with restriction enzymes BamH I and EcoR I, it was digested with the same enzyme double The digested vector pET32a(+) (purchased from Novagen, with His-tag c...

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Abstract

This invention discloses Gossypium hirsutum DREB transcription factor and its coding gene for application in breeding plants with improved stress resistance. The DREB transcription factor has one of the following amino acid residue sequences: (1) SEQ ID No. 1; (2) SEQ ID No. 1 with 1-10 amino acid residues substituted, deleted or added by proteins with transcriptional activation function. The DREB transcription factor and its coding gene can be applied in improving stress-resistance of plants.

Description

technical field [0001] The invention relates to a plant transcription factor and its encoding gene and application, in particular to a cotton-derived DREB transcription factor and its encoding gene, and its application in cultivating transgenic plants with improved stress resistance. Background technique [0002] Transcription factors, also known as trans-acting factors, are DNA-binding proteins that can specifically interact with cis-acting elements in the promoter region of eukaryotic genes. Through the interaction between them and other related proteins, they activate or inhibit transcription. The DNA binding region of a transcription factor determines its specificity for binding to cis-acting elements, while the transcriptional regulatory region determines whether it activates or inhibits gene expression. In addition, its own activity is also affected by nuclear localization and oligomerization. [0003] DREB transcription factors are transcription factors that can be ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/82
Inventor 刘进元黄波金龙国
Owner TSINGHUA UNIV
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