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Tulip glutathione S-transferase TfGST protein and encoding gene thereof

A technology of glutathione and transferase, applied in the field of plant molecular biology

Inactive Publication Date: 2014-04-16
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 glutathione S-transferase protein and its coding gene sequence

Method used

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  • Tulip glutathione S-transferase TfGST protein and encoding gene thereof
  • Tulip glutathione S-transferase TfGST protein and encoding gene thereof
  • Tulip glutathione S-transferase TfGST protein and encoding gene thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Embodiment 1, the cloning of tulip TfGST gene

[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 20220 No. 004) were planted and managed in the field according to conventional methods. Petal tissue was collected when the petals were fully colored for RNA extraction.

[0039] 2. Extraction of RNA

[0040] Total RNA was extracted using the "RNA prep pure plant total RNA extraction kit" (RNA prep pure Plant Kit: 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).

[0041] 3. Full-length cloning of genes

[0042] According to the conserved amino acid sequences of GST genes in other species, usin...

Embodiment 2

[0051] Example 2, Sequence Information and Homology Analysis of Tulip TfGST Gene

[0052] The full-length CDS open reading frame sequence of tulip TfGST is 663bp, and the detailed sequence is shown in SEQ ID NO.1; the amino acid sequence of tulip TfGST was deduced according to the CDS open reading frame sequence, with a total of 220 amino acid residues and a molecular weight of 24847.3 Dalton, the isoelectric point (pI) is 6.66, and the detailed sequence is shown in SEQ ID NO.2.

[0053] The CDS open reading frame sequence of tulip TfGST and the amino acid sequence of its encoded protein were nucleated 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 searches. Table 2 is the homologous comparison (GAP) result of the nucleotide sequence of tulip TfGST gene of the present invention and castor-oil plant (Ricinus communis) glutathione S-transferase gene mRN...

Embodiment 3

[0059] Embodiment 3, tulip TfGST gene expression difference in different developmental stages of flowers and in different tissues of tulip opposite sex

[0060] 1. Acquisition of materials

[0061] During the four 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 bulbs, aboveground stems, leaves and For the 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 a -80°C ultra-low temperature refrigerator for storage until use.

[0062] 2. Extraction of RNA

[0063] 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 flowers at different developmental stages and the RNA i...

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Abstract

The invention discloses a tulip glutathione S-transferase TfGST protein and an encoding gene thereof. The protein is a protein consisting of an ammonia acid sequence represented by SEQ ID NO.2 or a protein which is obtained by substitution, deficiency or adding of one or several ammonia acids on the ammonia acid sequence represented by SEQ ID NO.2 and has the activity of tulip glutathione S-transferase. The invention also provides a nucleotide sequence which is represented by SEQ ID NO.1 and is used for encoding the protein. A theoretical basis is supplied to regulation and control on the color of the a tulip by regulation and control on the spatio-temporal expression characteristic of a TfGST gene of the tulip through a gene engineering technology; the tulip glutathione S-transferase TfGST protein and the encoding gene have actual application value.

Description

technical field [0001] The invention belongs to the field of plant molecular biology, and relates to a key enzyme in the synthetic pathway of tulip anthocyanins and its coding gene, in particular to a tulip glutathione S-transferase TfGST protein and its coding gene. Background technique [0002] Anthocyanins are a class of flavonoids. In addition to imparting various colors to plant organs, anthocyanins play an important role in pollination, seed dispersal, protection from ultraviolet damage, and resistance to pathogen infection (Harborne and Williams2000; Wrolstad2004). Anthocyanins are synthesized through two processes, the phenylpropanoid pathway and the flavonoid pathway (Koses et al. 2005; Jeong et al. 2006). After formation, anthocyanins are toxic to cells due to their high biochemical activity and need to be transported from the cytoplasm to the vacuole for storage (Grotewold et al. 1998). Glutathione S-transferase (glutathione S-transferase, GST) can catalyze the c...

Claims

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Application Information

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IPC IPC(8): C12N9/10C12N15/54
CPCC12N9/1088C12Y205/01018
Inventor 袁媛史益敏唐东芹马晓红陶秀花
Owner SHANGHAI JIAO TONG UNIV
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