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Tulip TfMYB2 protein and coding gene thereof as well as probe

A coding and probe technology, applied in genetic engineering, plant genetic improvement, botanical equipment and methods, etc., can solve the problems of unclear MYB protein coding gene sequence and expression pattern, and no literature report.

Inactive Publication Date: 2014-04-16
SHANGHAI JIAOTONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The sequence and expression pattern of the MYB protein-encoding gene in tulip is still unclear, and there is no literature report related to the MYB protein and its encoding gene sequence in tulip

Method used

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  • Tulip TfMYB2 protein and coding gene thereof as well as probe
  • Tulip TfMYB2 protein and coding gene thereof as well as probe
  • Tulip TfMYB2 protein and coding gene thereof as well as probe

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1 , Tulip TfMYB2 gene cloning

[0039] 1. Acquisition of plant material

[0040] Healthy, uniform-sized tulip bulbs (Tulipa fosteriana 'Shangnong zaoxia', approved by the Shanghai Crop Variety Approval Committee. No.: Shanghai Nongpin Huahua 2011 No. 004) were planted as usual and managed in the field until the flowers were fully open , when the petals are fully colored, the petal tissue is collected for RNA extraction;

[0041] 2. Extraction of RNA

[0042] 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;

[0043] 3. Full-length cloning of genes

[0044] According to the amino acid conserved sequence of MYB protein in...

Embodiment 2

[0057] Example 2 , Sequence information and homology analysis of TfMYB2 gene in tulip

[0058] The full-length open reading frame sequence of the new tulip TfMYB2 gene of the present invention is 657bp, and its detailed sequence is shown in SEQ ID NO.3; according to the open reading frame sequence, the amino acid sequence of the tulip TfMYB2 protein is deduced, with a total of 218 amino acid residues. The molecular weight is 24851.8 Daltons, the isoelectric point (pI) is 8.25, and its detailed sequence is shown in SEQ ID NO.4;

[0059]The open reading frame sequence of the tulip TfMYB2 gene and the amino acid sequence of its encoded protein were nucleotide aligned in the GenBank+EMBL+DDBJ+PDB and Non-redundant GenBank CDS translations+PDB+SwissProt+Superdate+PIR databases using the BLAST program. Protein homology search, it was found that it has 77% identity with the lily LhMYB6 gene (GenBank accession number is AB534587.1) at the nucleotide level, such as figure 1 Shown (Q...

Embodiment 3

[0060] Example 3 , Tulip TfMYB2 gene expression differences

[0061] 1. Material acquisition: In 4 different developmental stages of flowers of 3 varieties of tulip (Tulipa fosteriana) with different flower colors (pink, light red, and bright red) (buds and petals are not colored; buds and petals are beginning to color; flower parts open, the petals are not fully colored; the flowers are fully open, the petals are completely colored), the petals are collected in the field, and the underground roots, aboveground stems, leaves and petals are collected at the same time (mixed samples of flower petals at various development stages), and the samples are respectively covered with aluminum platinum paper. Put it into liquid nitrogen immediately after wrapping, and then transfer it to a -80°C ultra-low temperature refrigerator for storage until use;

[0062] 2. RNA extraction: RNA prep pure Plant Total RNA Extraction Kit (RNA prep pure Plant Kit: Tiangen Biochemical Technology (Beij...

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Abstract

The invention provides a tulip TfMYB2 protein and a coding gene thereof as well as a probe. The protein is the protein in (a) or (b): (a) the protein comprising an amino acid sequence shown by SEQ ID NO.4; and (b) the protein which is derived from the protein in (a) through replacing, missing or adding one or more amino acids to the amino acid sequence shown by SEQ ID NO.4, and has the tulip MYB protein activity. The invention also provides a nucleotide sequence coding the protein, and the probe for detecting the nucleotide sequence. Expressions of a plurality of structural genes in biosynthetic pathways of flavonoid can be regulated and controlled through regulating the expression of TfMYB2 gene by utilizing the genetic engineering technology, or the TfMYB2 gene is transformed in other different plants, so that the content of flavonoid in transgenic plants can be effectively improved or lowered, the basis for achieving the purpose of flower color changing is provided, and the tulip TfMYB2 protein has important application value.

Description

technical field [0001] The present invention relates to a key enzyme in the synthetic pathway of tulip anthocyanins, its encoding gene and probe, and specifically relates to a tulip TfMYB2 protein, its encoding gene and its probe. Background technique [0002] Flowers are important reproductive organs of plants, and flower color is the prerequisite for normal reproduction of plants and also the important economic value of ornamental plants. Studying the molecular mechanism of flower color substance metabolism is the basis for breeding new flower varieties with different flower colors. The color of flowers is the result of the accumulation of anthocyanins. Anthocyanins mainly include flavonoids, carotenoids and betaines. The biosynthetic pathway of flavonoids is one of the most intensively studied metabolic pathways. [0003] The flavonoid biosynthetic pathway is mainly controlled by two classes of genes: structural genes and regulatory genes. Among them, structural genes...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/11C12Q1/68
Inventor 袁媛史益敏唐东芹马晓红
Owner SHANGHAI JIAOTONG UNIV
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