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Tulip chalcone isomerase TfCHI protein and coding gene thereof and probe

A technology of chalcone isomerase and tulip, applied in the direction of isomerase, genetic engineering, plant gene improvement, etc., can solve the problem of lack of tulip CHI protein

Inactive Publication Date: 2013-03-20
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 coding gene thereof and probe
  • Tulip chalcone isomerase TfCHI protein and coding gene thereof and probe
  • Tulip chalcone isomerase TfCHI protein and coding gene thereof and probe

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1 , Tulip TfCHI gene cloning

[0037] 1. Acquisition of plant material

[0038] 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;

[0039] 2. Extraction of RNA

[0040] 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 NANODROP 1000 Spectrophotometer) was used to determine the purity and concentration of RNA;

[0041] 3. Full-length cloning of genes

[0042] According to the amino acid conserved sequences ...

Embodiment 2

[0055] Example 2 , Sequence information and homology analysis of tulip TfCHI gene

[0056] 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;

[0057] 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 nucleoti...

Embodiment 3

[0058] Example 3 , the expression difference of tulip TfCHI gene in different developmental stages of flowers and in different tissues of tulip

[0059] 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;

[0060]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 a coding gene thereof and a probe. The protein is one selected from (a) and (b), wherein (a) is a protein composed of an amino acid sequence as represented by SEQ ID No. 4, and (b) is a protein derived from (a), obtained through replacement, deletion or addition of one or more amino acids of the amino acid sequence as represented by SEQ ID No. 4 and having activity of tulip chalcone isomerase. The invention also provides a nucleic acid sequence coding the protein and the probe for detection of the nucleic acid sequence. According to the invention, the tulip chalcone isomerase TfCHI protein and the coding gene thereof provide theoretical bases for changing flower colors and creating new flower colors through regulation and control of temporal-spatial expression characteristics of the tulip chalcone isomerase TfCHI protein by using gene engineering technology, and have a significant application value.

Description

technical field [0001] The invention relates to a key enzyme in the synthetic pathway of tulip anthocyanins, its encoding gene and probe, in particular to a tulip chalcone isomerase TfCHI protein, its encoding gene and its probe. Background technique [0002] The color of flowers is one of the important factors that determine their commodity value and ornamental value. The color of flowers is the result of the accumulation of anthocyanins, which mainly include flavonoids, carotenoids and betaines. As one of the three major anthocyanins, flavonoids give flowers a full range of colors from yellow to purple. The biosynthetic pathway of flavonoids is one of the more clearly studied metabolic pathways, and the biosynthetic pathway of anthocyanins is the most widely and deeply studied. Chalcone isomerase (CHI) is the second enzyme following chalcone synthase (CHS) in the anthocyanin biosynthetic pathway, which catalyzes the conversion of yellow chalcones to colorless flavanones ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/90C12N15/61C12Q1/68C12N15/11
CPCC12N9/90C12Y505/01006
Inventor 袁媛史益敏唐东芹马晓红
Owner SHANGHAI JIAO TONG UNIV
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