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Tulip chalcone isomerase TfCHI protein and encoding gene thereof

A technology of chalcone isomerase and tulip, which is applied in the field of biology and can solve problems such as the absence of tulip CHI protein

Inactive Publication Date: 2014-03-05
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 tulip CHI protein and its coding gene sequence

Method used

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  • Tulip chalcone isomerase TfCHI protein and encoding gene thereof
  • Tulip chalcone isomerase TfCHI protein and encoding gene thereof
  • Tulip chalcone isomerase TfCHI protein and encoding gene thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1 , the cloning of tulip TfCHI gene

[0041] 1. Acquisition of plant material

[0042] Healthy, uniform-sized tulip bulbs (Tulipa fosteriana 'Shangnongzaoxia', approved by the Shanghai Crop Variety Approval Committee. No.: Shanghai Nongpin Huahua 2011 No. 004) were planted and managed in the field according to the routine, and the flowers were fully opened. When the petals are fully colored, the petal tissue is collected for RNA extraction;

[0043] 2. Extraction of RNA

[0044] Use the "RNA prep pure Plant Total RNA Extraction Kit" to extract total RNA (RNA prep pure Plant Kit: Tiangen Biochemical Technology (Beijing) Co., Ltd.), use formaldehyde denaturing gel electrophoresis to identify the integrity of the RNA, and then analyze it in a spectrophotometer (Thermo Scientific NANODROP1000Spectrophotometer) was used to determine the purity and concentration of RNA;

[0045] 3. Full-length cloning of genes

[0046] According to the amino acid conserved seque...

Embodiment 2

[0059] Example 2 , Sequence information and homology analysis of tulip TfCHI gene

[0060]The full-length open reading frame sequence of the new tulip TfCHI gene of the present invention is 693bp, and the detailed sequence is shown in the sequence shown in SEQ ID NO.3. The amino acid sequence of tulip TfCHI was deduced according to the sequence of the open reading frame, with a total of 230 amino acid residues, a molecular weight of 24374.9 Daltons, and an isoelectric point (pI) of 5.15. For the detailed sequence, see the sequence shown in SEQ ID NO.4;

[0061] The open reading frame sequence of tulip TfCHI and the amino acid sequence of its encoded protein were analyzed by BLAST program in the databases of Non-redundant GenBank+EMBL+DDBJ+PDB and Non-redundant GenBank CDS translations+PDB+SwissProt+Superdate+PIR acid and protein homology search, it was found that it has 75% identity with the American oil palm CHI gene (GenBank accession number is FJ940770.1) at the nucleotid...

Embodiment 3

[0062] Example 3 , the expression difference of tulip TfCHI gene in different developmental stages of flowers and in different tissues of tulip

[0063] 1. Obtaining materials: During the four different development stages of tulip flowers (buds, petals are not colored; buds, petals are beginning to be colored; flowers are partially open, petals are not fully colored; flowers are fully open, petals are fully colored), and other tulips are collected in the field. For the petals, the leaves, aboveground stems, stamens, pistils, and petals (the mixed samples of petals at each coloring stage) were collected at the same time, and the samples were wrapped in aluminum platinum paper and immediately put into liquid nitrogen, and then transferred to -80°C ultra-low temperature Store in the refrigerator for later use;

[0064] 2. RNA extraction: RNA prep pure Plant Total RNA Extraction Kit (RNA prep pure Plant Kit: Tiangen Biochemical Technology (Beijing) Co., Ltd.) was used to extract...

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Abstract

The invention relates to a tulip chalcone isomerase TfCHI protein and an encoding gene thereof. The protein is composed of amino acid sequences represented by the SEQ ID No.4, or the protein is composed of amino acid sequences represented by the SEQ ID No.4 which have been processed by replacing, removing, or adding one or several amino acids, and has a tulip chalcone isomerase activity. The invention also provides a nucleic acid sequence which encodes the protein mentioned above and is represented by the SEQ ID No.3. By utilizing the tulip chalcone isomerase gene provided by the invention and various screening method, substances, acceptors, inhibitors or antagonists which can interact with the chalcone isomerase TfCHI can be screened out. By utilizing the tulip chalcone isomerase gene provided by the invention, people can change the flower color through a gene engineering method, and a novel way is provided for creating new flower colors.

Description

technical field [0001] The invention relates to a key enzyme and its encoding gene in the synthesis pathway of tulip anthocyanin, in particular to a tulip chalcone isomerase TfCHI protein and its encoding gene, and belongs to the technical field of biology. Background technique [0002] Chalcone isomerase (CHI) is the second enzyme following chalcone synthase (CHS) in the flavonoid synthesis pathway. Under the catalysis of CHI, a chalcone molecule undergoes an intramolecular reaction The C-ring is closed to convert to flavanones. An important feature of this reaction is the transformation of yellow chalcones into colorless flavanones, which can be used as an important biomarker for studying the function of CHI. Almost all flavonoids in plants are derived from flavanones. Chalcone isomerase is the key enzyme to increase flavonoids. Flavonoids are a class of polyphenolic compounds with similar structures. Most of them have color and play an important role in plant growth and...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/90C12N15/61
CPCC12N9/90C12Y505/01006
Inventor 袁媛史益敏唐东芹马晓红
Owner SHANGHAI JIAO TONG UNIV
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