Tulip flavonoid-3'-hydroxylase TfF3' H protein and its coding gene
A technology of flavonoids and tulips, applied in the molecular field, can solve problems such as the unknown
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Examples
Embodiment 1
[0051] Example 1 , the cloning of tulip TfF3'H gene
[0052] 1. Acquisition of plant material
[0053] 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.
[0054] 2. Extraction of RNA
[0055] 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).
[0056] 3. Full-length cloning of the gene
[0057] According to the amino acid conserved sequence of F3'H ...
Embodiment 2
[0071] Example 2 , Sequence information and homology analysis of tulip TfF3'H gene
[0072] The full-length CDS open reading frame sequence of tulip TfF3'H is 1533bp, and the detailed sequence is shown in the sequence shown in SEQ ID NO.3; the amino acid sequence of tulip TfF3'H is deduced according to the CDS open reading frame sequence, with a total of 510 amino acid residues , with a molecular weight of 55482.2 Daltons and an isoelectric point (pI) of 6.98. See the sequence shown in SEQ ID NO.4 for the detailed sequence.
[0073] The CDS open reading frame sequence of tulip TfF3'H and the amino acid sequence of its encoded protein were stored 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 were carried out, and it was found that it has 82% identity with the lily F3'H gene (GenBank accession number AB699162.1) at the nucleotide level,...
Embodiment 3
[0074] Example 3 , Expression differences of tulip F3'H gene in different developmental stages of flowers and in different tissues of tulip
[0075] 1. Acquisition of materials
[0076] 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.
[0077] 2. Extraction of RNA
[0078] 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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Application Information
- IPC
- C12N9/02; C12N15/53
- CPC
- C12N9/0073; C12Y114/13021
- Inventors
- 袁媛; 史益敏