The auxin response factor aparf2 of Agapanthus chinensis and its coding gene and probe

A plant auxin and coding technology, which is applied in the field of Agapanthus Agapanthus auxin response factor ApARF2 and its coding gene and probe, can solve the problems of no coding gene sequence, etc., and achieve high ornamental value, wide application, and great application value Effect

Inactive Publication Date: 2019-06-21
SHANGHAI JIAOTONG 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 ARF protein and its coding gene sequence

Method used

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  • The auxin response factor aparf2 of Agapanthus chinensis and its coding gene and probe
  • The auxin response factor aparf2 of Agapanthus chinensis and its coding gene and probe
  • The auxin response factor aparf2 of Agapanthus chinensis and its coding gene and probe

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Embodiment 1, the cloning of Agapanthus Abaciens ApARF2 gene

[0045] 1. Acquisition of plant material

[0046] Agapanthus praecox ssp.Orientalis (Agapanthus praecox ssp.Orientalis)(Zhang D.,Zhuo L.H.,ShenX.H.Sporogenesis and gametogenesis in Agapanthus praecox Willd.orientalis(Leighton)Leighton and their systematic implications.Plant Syst.Evol.2010,288:1 -11.) Adult seedling leaf tissue for extracting RNA;

[0047] 2. Extraction of RNA

[0048] Use "RNA prep pure plant total RNA extraction kit" to extract total RNA (Trizol: Invitrogen), use formaldehyde denaturing gel electrophoresis to identify the integrity of RNA, and then measure the purity and concentration of RNA on a spectrophotometer (Thermo Scientific NANODROP1000Spectrophotometer) ;

[0049] 3. Full-length cloning of genes

[0050] According to the protein function annotation results of Agapanthus japonica transcriptome sequencing (RNA-seq), the core fragment of ApARF2 gene of Agapanthus japonica was ob...

Embodiment 2

[0073] Example 2, Sequence Information and Homology Analysis of Agapanthus Abaciana ApARF2 Gene

[0074]The full-length open reading frame sequence of the new Agapanthus agapanthus ApARF2 gene of the present invention is 2349bp, 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 the Agapanthus Abaciens ApARF2 protein is deduced, with a total of 782 amino acid residues, a molecular weight of 87.78kDa, and an isoelectric point (pI) of 6.36. For the detailed sequence, see the sequence shown in SEQ ID NO.4;

[0075] The open reading frame sequence of Agapanthus abaciens ApARF2 and the amino acid sequence of its encoded protein were nuclearized in Non-redundant GenBank+EMBL+DDBJ+PDB and Non-redundant GenBank CDS translations+PDB+SwissProt+Superdate+PIR databases using BLAST program Nucleotide and protein homology search, it was found that it has 91% identity with the mulberry ARF gene (...

Embodiment 3

[0076] Embodiment 3, model plant Arabidopsis thaliana transformed with Agapanthus ApARF2 gene

[0077] 1. Construction of the expression vector containing the target gene (Agapanthus Abaciens ApARF2 gene)

[0078] According to the full-length coding sequence of Agapanthus ApARF2 gene (SEQ ID NO.3), design primers to amplify the complete coding reading frame, and introduce restriction endonuclease sites on the upstream and downstream primers respectively (this can be determined by the selected vector Depends), in order to construct the expression vector. Using the amplified product obtained in Example 1 as a template, after PCR amplification, the coding region sequence of the Agapanthus Agapanthus ApARF2 gene was connected to an intermediate vector (such as pMD19-T) for sequencing, and then the sequenced correct Agapanthus ApARF2 The coding region sequence of the gene is further cloned into an expression vector (such as pHB), and it is transformed into Agrobacterium tumefaci...

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Abstract

The invention relates to an African agapanthus auxin response factor ApARF2 and an encoding gene and probe thereof. Protein is shown in a or b, wherein a is protein formed by an amino acid sequence shown in SEQ ID NO.4, and b is protein which is formed by substitution or deletion or addition of one or more amino acids to the amino acid sequence shown in SEQ ID NO.4, has African agapanthus ApARF2 protein activity and is derived from a. The invention further provides a nucleotide sequence for encoding the protein and the probe for detecting the nucleotide sequence. An African agapanthus auxin signal transduction pathway is adjusted and controlled through the gene engineering technology, the purposes of controlling growth and development of African agapanthus and establishing organ morphology are achieved accordingly, a theoretical basis is provided for molecular breeding, and high application value is achieved.

Description

technical field [0001] The present invention relates to Agapanthia auxin response factor ApARF2 and its encoding gene and probe, and specifically relates to Agapanthia auxin response factor ApARF2 and its encoding gene and probe. Background technique [0002] 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. Among them, the auxin response factor (ARF) is a kind of transcription factor that regulates the expression of auxin-responsive genes. It is at the center of the auxin signal transduction process. The auxin response element in the promoter region of the response gene specifical...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/82A01H5/00A01H6/20C12Q1/6895C12N15/11
CPCC07K14/415C12N15/8294C12Q1/6895
Inventor 陈冠群申晓辉杨舟陈淑敏张荻
Owner SHANGHAI JIAOTONG UNIV
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