Tulipa gesneriana TfbHLH1 protein, encoding gene thereof and probe

A coding and protein technology, applied in the fields of biochemical equipment and methods, enzymes, DNA/RNA fragments, etc., can solve problems such as no literature report, unclear bHLH protein coding gene sequence and expression pattern, etc.

Inactive Publication Date: 2015-01-14
SHANGHAI JIAOTONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The gene sequence and expression pattern of the bHLH protein in tulip is not clear, and there is no literature report related to the bHLH protein and its coding gene sequence in tulip

Method used

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  • Tulipa gesneriana TfbHLH1 protein, encoding gene thereof and probe
  • Tulipa gesneriana TfbHLH1 protein, encoding gene thereof and probe
  • Tulipa gesneriana TfbHLH1 protein, encoding gene thereof and probe

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1 , Tulip TfbHLH1 gene cloning

[0042] 1. Acquisition of plant material

[0043] Healthy, uniform-sized tulip bulbs (Tulipa fosteriana 'Shangnong zaoxia', 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 , when the petals are fully colored, the petal tissue is collected for RNA extraction;

[0044] 2. Extraction of RNA

[0045] Use "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 RNA, and then analyze the RNA in a spectrophotometer (Thermo Scientific NANODROP1000Spectrophotometer) was used to measure the purity and concentration of RNA;

[0046] 3. Full-length cloning of genes

[0047] According to the conserved amino acid sequences of bHLH proteins ...

Embodiment 2

[0060] Example 2 , Sequence Information and Homology Analysis of Tulip TfbHLH1 Gene

[0061]The full-length open reading frame sequence of the new tulip TfbHLH1 gene of the present invention is 2079bp, and the detailed sequence is shown in SEQ ID NO.3. According to the open reading frame sequence, the amino acid sequence of the tulip TfbHLH1 protein is deduced, with a total of 692 amino acid residues. The molecular weight is 77655.3 Daltons, and the isoelectric point (pI) is 4.87. For the detailed sequence, see the sequence shown in SEQ ID NO.4;

[0062] The open reading frame sequence of the tulip TfbHLH1 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 76% identity with the lily LhbHLH1 gene (GenBank accession number is AB222075.1) at the nucleotide level, as ...

Embodiment 3

[0063] Example 3 , Expression differences of tulip TfbHLH1 gene in different tissues and different developmental stages of flowers

[0064] 1. Material acquisition: in 4 different developmental stages of tulip (Tulipa fosteriana 'Shangnong zaoxia') flowers (buds, petals uncolored; buds, petals start to color; flowers partially open, petals not fully colored; flowers fully open, petals completely colored), take its petals in the field, and take the aboveground stems, leaves, stamens, pistils and petals (mixed samples of petals at each development stage) at the same time, wrap the samples with aluminum platinum paper, put them into liquid nitrogen immediately, and then transfer them to Store in a -80°C ultra-low temperature refrigerator for use;

[0065] 2. Extraction of RNA: 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 provides a tulipa gesneriana TfbHLH1 protein, encoding gene thereof and a probe. The protein is a protein (a) or (b): (a), the protein is composed of an amino acid sequence shown in SEQ ID NO. 4; and (b), the protein derived from (a) with the activity of the protein of tulipa gesneriana bHLH, and obtained through performing replacement and / or deletion and / or addition of one or more amino acids for the amino acid sequence shown in SEQ ID NO. 4. The invention further provides a nucleotide sequence for encoding the protein and a probe for detecting the nucleotide sequence. According to the invention, the genetic engineering technology is utilized, the flavonoid content is effectively increased or reduced and a foundation is provided for color change through adjusting the transcriptional level of a plurality of structural genes in the expression regulation flavonoid biosynthetic pathway of the TfbHLH1 gene, and important theoretical and practical application value can be obtained.

Description

technical field [0001] The present invention relates to a key enzyme in the synthetic pathway of tulip anthocyanins, its encoding gene and probe, in particular to a tulip TfbHLH1 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 directly ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/00C12N15/52C12N15/11
Inventor 袁媛史益敏唐东芹马晓红
Owner SHANGHAI JIAOTONG UNIV
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