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Agapanthus praecox auxin receptor protein TIR1 and coding gene and probe thereof

A technology of receptor protein and auxin, which can be used in genetic engineering, plant genetic improvement, biochemical equipment and methods, etc., and can solve problems that have not yet been solved

Inactive Publication Date: 2013-10-02
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no literature report related to Agapanthus TIR1 protein and its coding gene sequence

Method used

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  • Agapanthus praecox auxin receptor protein TIR1 and coding gene and probe thereof
  • Agapanthus praecox auxin receptor protein TIR1 and coding gene and probe thereof
  • Agapanthus praecox auxin receptor protein TIR1 and coding gene and probe thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1 , Cloning of Agapanthus TIR1 gene

[0041] 1. Acquisition of plant material

[0042] The leaf tissue of Agapanthus adult seedlings was taken for RNA extraction; Agapanthus was introduced into my country from South Africa in 2000, and the introduction experiment was carried out in Shanghai. In his master's thesis "Research on the Dwarfing of Agapanthus Potted Plants" published in 2011, Sun Bo introduced the basic biological characteristics and introduction history of Agapanthus in detail. The agapanthus material involved in this embodiment can be obtained through public channels.

[0043] 2. Extraction of RNA

[0044] Total RNA (Trizol: Invitrogen) was extracted with "RNA prep pure plant total RNA extraction kit", the integrity of RNA was identified by formaldehyde denaturing gel electrophoresis, and then the purity and concentration of RNA were determined on a spectrophotometer (Thermo Scientific NANODROP1000Spectrophotometer) ;

[0045] 3. Full-length c...

Embodiment 2

[0066] Example 2 , Sequence information and homology analysis of Agapanthus TIR1 gene

[0067] The full-length open reading frame sequence of the novel Agapanthus TIR1 gene of the present invention is 1767bp, and the detailed sequence is shown in the sequence shown in SEQ ID NO.3. According to the open reading frame sequence, the amino acid sequence of Agapanthus TIR1 protein was deduced, with a total of 588 amino acid residues, a molecular weight of 65.8kDa, and an isoelectric point (pI) of 5.83. For the detailed sequence, see the sequence shown in SEQ ID NO.4;

[0068] The open reading frame sequence of Agapanthus TIR1 and the amino acid sequence of its encoded protein were nucleated in Non-redundant GenBank+EMBL+DDBJ+PDB and Non-redundant GenBank CDS translations +PDB+SwissProt+Superdate+PIR databases using BLAST program Nucleic acid and protein homology search, and found that it has 72% identity with castor plant TIR1 gene (accession number: XM_002524640.1) at the nucleo...

Embodiment 3

[0069] Example 3 , Expression differences of Agapanthus TIR1 gene at different developmental stages of somatic embryos and under exogenous regulation

[0070] 1. Acquisition of materials: samples were taken from four different developmental stages of Agapanthus somatic embryos (callus; embryogenic callus; somatic embryos; somatic embryo seedlings); The 3-year-old plants of Agapanthus basilica regulate auxin signal. The exogenous regulatory substances are IAA (100ppm) and NPA (auxin polar transport inhibitor: 100ppm). After treatment, the control sample (no treatment), IAA treatment and NPA Leaves of treated plants were sampled. Wrap the samples with aluminum platinum paper and put them into liquid nitrogen immediately, and then transfer them to -80°C ultra-low temperature refrigerator for storage until use;

[0071] 2. RNA extraction: use RNA prep pure plant total RNA extraction (Trizol: Invitrogen); extract total RNA from different sample tissues of Agapanthus lily;

[00...

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Abstract

The invention relates to an agapanthus praecox auxin receptor protein TIR1 and a coding gene and probe thereof. The protein is: (a) a protein consisting of the amino acid sequence shown in SEQ ID No.4; or (b) a protein obtained by substituting, losing or adding one or multiple amino acids of the amino acid sequence shown in SEQ ID No.4, having agapanthus praecox auxin receptor protein activity and derived from (a). The invention also provides a nucleotide sequence for coding the protein and a probe for detecting the nucleotide sequence. According to the agapanthus praecox auxin receptor protein TIR1 and the coding gene and probe thereof provided by the invention, the agapanthus praecox auxin signal transduction pathway is regulated by the genetic engineering technology to reach an aim of controlling the growth development and organ morphogenesis, thereby providing a theoretical basis for molecular breeding and creating great application value.

Description

technical field [0001] The present invention relates to the key protein TIR1 and its coding gene and probe in the Agapanthus abacifolia auxin signal transduction pathway, and specifically relates to an Agapanthus agapanthus auxin receptor protein TIR1 and its coding gene and probe. Background technique [0002] Endogenous hormones play an important role in the growth and development of ornamental plants and the regulation of ornamental traits. Auxin is the only endogenous hormone with polar transport characteristics in plants. The content and distribution of auxin are related to the growth and development speed of plants and the polar structure and spatial form of various organs and tissues. The signal transduction pathway of auxin is relatively clear at present, in which auxin must bind to its receptor protein to regulate the downstream gene network and cause physiological responses, and TIR1 protein is the most important auxin receptor protein. [0003] Somatic embryogene...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12Q1/68C12N15/11
Inventor 申晓辉张荻岳建华任丽
Owner SHANGHAI JIAO TONG UNIV
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