The auxin response factor aparf1 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 ApARF1 and its coding gene and probe, can solve the problems of no coding gene sequence, etc., and achieve high ornamental value, great application value and wide application Effect

Active 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 aparf1 of Agapanthus chinensis and its coding gene and probe
  • The auxin response factor aparf1 of Agapanthus chinensis and its coding gene and probe
  • The auxin response factor aparf1 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 ApARF1 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 ApARF1 gene of Agapanthus japonica was ob...

Embodiment 2

[0073] Example 2, Sequence Information and Homology Analysis of Agapanthus Abaciens ApARF1 Gene

[0074]The full-length open reading frame sequence of the new Agapanthus agapanthus ApARF1 gene of the present invention is 1770bp, 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 ApARF1 protein is deduced, with a total of 589 amino acid residues, a molecular weight of 65.99kDa, 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 abaculata ApARF1 and the amino acid sequence of its encoded protein were nuclearized in the Non-redundant GenBank+EMBL+DDBJ+PDB and Non-redundant GenBank CDS translations+PDB+SwissProt+Superdate+PIR databases using the BLAST program Nucleotide and protein homology search, it was found that it has 98% identity with Nicotiana taba...

Embodiment 3

[0076] Embodiment 3, model plant Arabidopsis thaliana transformed with Agapanthus ApARF1 gene

[0077] 1. Construction of the expression vector containing the target gene (Agapanthus Abaciens ApARF1 gene)

[0078] According to the full-length coding sequence of Agapanthus ApARF1 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 sequence of the coding region of the Agapanthus Agapanthus ApARF1 gene was connected to an intermediate vector (such as pMD19-T) for sequencing, and then the sequenced correct Agapanthus ApARF1 The coding region sequence of the gene is further cloned into an expression vector (such as pHB), and it is transformed into Agrobacterium t...

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Abstract

The invention discloses an agapanthus auxin response factor ApARF1 and an encoding gene and a probe thereof. A protein is a protein shown in (a) or (b): (a) a protein formed by an amino acid sequence shown in SEQ ID NO.4, and (b) a protein which is formed by replacement or deletion or addition of one or more amino acids on the amino acid sequence shown in SEQ ID NO.4, has agapanthus ApARF1 protein activity and is derived from (a). The invention further provides a nucleotide sequence for encoding the protein and a probe for detecting the nucleotide sequence. An agapanthus auxin signal transcriptional pathway is controlled through a gene engineering technology, so that the purposes of controlling growth and development of agapanthus and organ morphology establishment are achieved, the theoretical basis is provided for molecular breeding, and high application value is achieved.

Description

technical field [0001] The present invention relates to Agapanthus auxin response factor ApARF1 and its encoding gene and probe, and specifically relates to Agapanthia auxin response factor ApARF1 and its encoding gene and probe. Background technique [0002] Auxin is an important growth regulator molecule in plants, which is involved in the growth and development of plant roots and stems, the aging of organs, the formation and differentiation of vascular tissue, the maintenance of apical dominance, the establishment of the embryonic axis, and the plant's orientation and orientation. Many processes of growth and development, such as light response and stimulation of floral organ growth, play an important role in the whole life cycle of plants. Auxin response factor (ARF) is a class of transcription factors that regulate the expression of auxin-responsive genes. It plays a central role in the process of auxin signal transduction. It can specifically bind to auxin-responsive e...

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 张荻杨舟陈冠群张敏涛张莹
Owner SHANGHAI JIAOTONG UNIV
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