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Tulip chalcone synthase tfchs protein and its coding gene

A chalcone synthase, tulip technology, applied in genetic engineering, plant genetic improvement, enzymes and other directions, can solve problems such as the absence of tulip CHS protein

Inactive Publication Date: 2016-09-07
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 tulip CHS protein and its coding gene

Method used

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  • Tulip chalcone synthase tfchs protein and its coding gene
  • Tulip chalcone synthase tfchs protein and its coding gene
  • Tulip chalcone synthase tfchs protein and its coding gene

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1 , Tulip TfCHS gene cloning

[0046] 1. Acquisition of plant material

[0047] 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. Petal tissue was collected when the petals were fully colored for RNA extraction.

[0048] 2. Extraction of RNA

[0049] Total RNA was extracted with "RNA prep pure Plant Total RNA Extraction Kit" (RNA prep pure PlantKit: Tiangen Biochemical Technology (Beijing) Co., Ltd.). 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 NANODROP1000 Spectrophotometer).

[0050] 3. Full-length cloning of genes

[0051] According to the conserved amino acid sequence of the CHS gene, using th...

Embodiment 2

[0062] Example 2 , Sequence information and homology analysis of the TfCHS gene of tulip

[0063] The full-length CDS open reading frame sequence of tulip TfCHS is 1179bp. For the detailed sequence, see the sequence shown in SEQ ID NO.3. According to the CDS open reading frame sequence, the amino acid sequence of tulip TfCHS is deduced, with a total of 392 amino acid residues and a molecular weight of 42861.3 lanes It has an isoelectric point (pI) of 5.90, and its detailed sequence is shown in SEQ ID NO.4.

[0064] The CDS open reading frame sequence of tulip TfCHS and the amino acid sequence of its encoded protein were analyzed by BLAST program in Non-redundant GenBank+EMBL+DDBJ+PDB and Non-redundant GenBank CDS translations+PDB+SwissProt+Superdate+PIR database Acid and protein homology search, it was found that it has 86% identity with lily CHS gene (GenBank accession number HQ161731.1) at the nucleotide level, such as figure 1 and figure 2 shown; at the amino acid lev...

Embodiment 3

[0065] Example 3 , Expression difference of tulip TfCHS gene in different developmental stages of flowers and in different tissues of tulip

[0066] 1. Acquisition of materials

[0067] In the 4 different developmental stages of tulip flowers (buds, petals are not colored; buds, petals are beginning to color; flowers are partially open, petals are not fully colored; flowers are fully open, petals are fully colored), the petals are collected in the field, and the leaves are collected at the same time , aerial stems, stamens, pistils, and petals (mixed samples of petals at each coloring stage), the samples were wrapped in aluminum platinum paper and immediately put into liquid nitrogen, and then transferred to -80°C ultra-low temperature refrigerator for storage until use.

[0068] 2. Extraction of RNA

[0069] RNA prep pure Plant Total RNA Extraction Kit (RNA prep pure Plant Kit: Tiangen Biochemical Technology (Beijing) Co., Ltd.) was used to extract the petals of tulip flow...

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Abstract

The invention relates to a tulip chalcone synthase TfCHS protein, and a coding gene and a probe thereof. The protein is (a) or (b) as follows: (a) a protein composed of an amino acid sequence shown as a SEQ ID NO.4; and (b) a protein obtained by substitution, deletion or addition of one or several amino acids on the amino acid sequence shown as the SEQ ID NO.4, having tulip chalcone synthase activity and derived from (a). The invention also provides a nucleic acid sequence encoding the protein, and the probe to detect the above nucleic acid sequence. The invention provides theoretical basis for regulation of spatio-temporal expression characteristic of the TfCHS gene of tulip for color change and color innovation by using a genetic engineering technology, and has important application value.

Description

technical field [0001] The invention relates to a key enzyme and its coding gene in the synthesis pathway of tulip anthocyanin, in particular to a tulip chalcone synthase TfCHS protein and its coding gene, and belongs to the field of biotechnology. Background technique [0002] The color of flowers is one of the important factors that determine the commodity value and ornamental value of flowers. Changing and innovating flower colors can increase the commodity value and ornamental value of flowers. The color of flowers is the result of the accumulation of anthocyanins in petal cells, mainly including flavonoids, carotenoids and betaines. Flavonoids are a large class of anthocyanins that give flowers their color from yellow to purple. The biosynthetic pathway of flavonoids can be divided into three stages, the first stage is from phenylalanine to coumaryl-CoA, which is common to many secondary metabolisms; the second stage is from coumaryl-CoA to dihydroflavonol , which is...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/10C12N15/54
CPCC12N9/1037C12Y203/01074
Inventor 袁媛史益敏唐东芹马晓红
Owner SHANGHAI JIAOTONG UNIV
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