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Tulip flavonoid 3-O-glucosyltransferase Tf3GT protein and coding gene thereof

A glucosyl and flavonoid technology, applied in the field of molecular biology, can solve problems such as

Inactive Publication Date: 2014-04-02
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 UF3GT protein and its coding gene sequence

Method used

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  • Tulip flavonoid 3-O-glucosyltransferase Tf3GT protein and coding gene thereof
  • Tulip flavonoid 3-O-glucosyltransferase Tf3GT protein and coding gene thereof
  • Tulip flavonoid 3-O-glucosyltransferase Tf3GT protein and coding gene thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] The cloning of embodiment 1, tulip Tf3GT gene

[0047] 1. Acquisition of plant material

[0048] Healthy, uniform-sized tulip bulbs (Tulipa fosteriana 'Shangnongzaoxia', approved by the Shanghai Crop Variety Approval Committee. Number: Shanghai Nongpin Huahua 2011 No. 004) were planted as usual and managed in the field until the flowers were fully open. Petal tissue was collected when the petals were fully colored for RNA extraction.

[0049] 2. Extraction of RNA

[0050] 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 NANODROP 1000 Spectrophotometer).

[0051] 3. Full-length cloning of genes

[0052] According to the conserved amino acid sequences of FLS genes in other ...

Embodiment 2

[0061] Example 2, Sequence Information and Homology Analysis of Tulip Tf3GT Gene

[0062] The full-length CDS open reading frame sequence of tulip Tf3GT is 1371bp, and the detailed sequence is shown in SEQ ID NO.1; the amino acid sequence of tulip Tf3GT is deduced according to the CDS open reading frame sequence, with a total of 456 amino acid residues and a molecular weight of 48492 Dalton, the isoelectric point (pI) is 6.02, and the detailed sequence is shown in SEQ ID NO.2.

[0063] The CDS open reading frame sequence of tulip Tf3GT and the amino acid sequence of its encoded protein were nucleated 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 searches. Table 2 is the result of the homology comparison (GAP) of the nucleotide sequence of the tulip Tf3GT gene of the present invention and the nucleotide sequence of the Dutch iris (Iris holland...

Embodiment 3

[0069] Embodiment 3, tulip Tf3GT gene expression difference in different developmental stages of flowers and in different tissues of tulip opposite sex

[0070] 1. Acquisition of materials

[0071] 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), Yu Tianwen collected its bulbs, ground stems, leaves As 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 -80°C ultra-low temperature refrigerator for storage until use.

[0072] 2. Extraction of RNA

[0073] 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 ...

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Abstract

The invention discloses a tulip flavonoid 3-O-glucosyltransferase Tf3GT protein and a coding gene thereof. The protein is a protein composed of amino acid sequence disclosed as SEQ ID NO.2, or a protein with tulip flavonoid 3-O-glucosyltransferase activity obtained by substitution, deletion or addition of one or more amino acids on the basis of the amino acid sequence disclosed as SEQ ID NO.2. The invention also provides a nucleic acid sequence disclosed as SEQ ID NO.1 for coding the protein. When the tulip flavonoid 3-O-glucosyltransferase Tf3GT gene is expressed in Escherichia coli cells, the recombinant flavonoid 3-O-glucosyltransferase protein can promote anthocyanin and UDP-glucose to react and generate the anthocyanin. The Tf3GT coding sequence has application value in modification of varieties by a transgenic technique.

Description

technical field [0001] The invention belongs to the field of molecular biology, in particular to a tulip flavonoid 3-O-glucosyltransferase Tf3GT protein and its coding gene. Background technique [0002] In the process of plant flower color formation, UDP-glucose: flavonoid 3-O-glucosyltransferase (UDP-glucose: flavonoid 3-O-glucosyltransferase, UF3GT, EC 2.4.1.91) catalyzes the transfer of UDP-glucose to anthocyanin C The C3 hydroxyl position of the ring (Carolye EL, 1996; Masako FM, 2003), converts colorless unstable anthocyanins into colored stable anthocyanins. Anthocyanins exist in vacuoles in a water-soluble form, so UF3GT is considered an indispensable enzyme in the anthocyanin synthesis pathway. Glycosylation modification can improve the stability of anthocyanins on the one hand, and affect the anthocyanin synthesis on the other hand. Plain color changes. For example, a large amount of expression of UF3GT gene can be detected in red grape peel, while UF3GT gene is ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/10C12N15/54C12Q1/68C12N15/11C12N15/70
CPCC12N9/1051C12Y204/01091
Inventor 袁媛史益敏唐东芹马晓红陶秀花
Owner SHANGHAI JIAO TONG UNIV
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