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Scutellaria viscidula chalcone synthetase albumen coded sequence

A technology of chalcone and coding sequence, which is applied in the field of genetic engineering and can solve problems such as the cloning of the chs gene of Scutellaria baicalensis, which has not been discovered or seen.

Inactive Publication Date: 2008-08-13
SOUTHWEST UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In the analysis of existing documents, there is no cloning of the chs gene on the biosynthetic pathway of Scutellaria baicalensis flavonoids, and no reports of documents closely related to the subject of the present invention have been found so far

Method used

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  • Scutellaria viscidula chalcone synthetase albumen coded sequence
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  • Scutellaria viscidula chalcone synthetase albumen coded sequence

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Embodiment 1

[0055] Cloning of chalcone synthase from Scutellaria baicalensis

[0056] 1. Tissue separation (isolation)

[0057] Scutellaria baicalensis was purchased from Shanxi University, and the seeds were sown in the greenhouse. When the Scutellaria baicalensis seedlings grew to a height of 10 cm, DNA or RNA was prepared to be extracted.

[0058] 2. RNA isolation (RNA isolation)

[0059] Take part of the tissue, grind it with a mortar, add it to a 1.5mL EP tube filled with lysate, shake it fully, and then transfer it into a glass homogenizer. After homogenization, transfer to 1.5mL EP tube, and extract total RNA (TRIzol Reagents, GIBCO BRL, USA). The quality of total RNA was identified by formaldehyde denaturing gel electrophoresis, and then the RNA content was determined on a spectrophotometer.

[0060] 3. Cloning of Full-length cDNA

[0061] According to the conserved nucleotide sequences of chalcone synthase genes of various plants, using the principle of homologous gene clonin...

Embodiment 2

[0072] Chalcone synthase gene sequence information and homology analysis of Scutellaria baicalensis

[0073] The full-length cDNA of the new chalcone synthase gene sequence of the present invention is 1142bp in length, and the detailed sequence is shown in SEQ ID NO.3, wherein the open reading frame is located at 54-752 nucleotides (699 nucleotides). According to the full-length cDNA, the amino acid sequence of chalcone synthase from Scutellaria baicalensis was deduced, which has a total of 233 amino acid residues, a molecular weight of 24.78kDa, and an isoelectric point (pI) of 4.80. See SEQ ID NO.4 for the detailed sequence.

[0074] The full-length cDNA sequence related to the chalcone synthase gene gene of Scutellaria baicalensis and its encoded protein were stored in the Non-redundant GenBank+EMBL+DDBJ+PDB and Non-redundant GenBankCDS translations+PDB+SwissProt+Superdate+PIR databases using the BLAST program Nucleotide and protein homology searches were carried out, and ...

Embodiment 3

[0076] Eukaryotic cell expression of chalcone synthase protein or polypeptide in Scutellaria baicalensis and detection of baicalin content in transgenic plants.

[0077] The construction of the expression vector containing the target gene, according to the Scutellaria baicalensis chalcone synthase gene coding sequence (SEQ ID NO.3), design and amplify the primers for the complete coding reading frame, and introduce restriction on the upstream and downstream primers respectively Endonuclease sites (this may depend on the carrier selected), in order to construct an expression vector. Using the amplified product obtained in Example 1 as a template, after PCR amplification, the Scutellaria baicalensis chalcone synthase gene cDNA was cloned into an intermediate vector (such as pBluescript), and further cloned into a binary expression vector (such as pBI121 and Improved pCAMBIA1304), the expression vector was identified under the premise of ensuring the correct reading frame, and th...

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Abstract

A collecting hairs baikal skullcap root chalcone ketone synthetase albumen coding sequence belongs to genetic engineering field comprises encoding polypeptides nucleotide sequence possessing chalcone ketone synthetase albumen activity which the nucleotide sequence and the 54-752 bit nucleotide sequence in SEQ ID NO.3 has 70124564mology, or the nucleotide sequence can cross-fertilize with the 54-752 bit nucleotide sequence in SEQ ID NO.3 in condition of 45-55 DEG C. The invention has evident action for improving the content of flavonoid such as baikal skullcap root glycosides in collecting hairs baikal skullcap root plant material also the invention has prodigious applications worthiness.

Description

technical field [0001] The invention relates to a protein coding sequence, in particular to a chalcone synthase protein coding sequence of Scutellaria baicalensis, which belongs to the field of genetic engineering. Background technique [0002] Scutellaria baicalensis is a perennial medicinal herb belonging to the genus Scutellariae of the Lamiaceae. There are more than 300 species of plants in this genus, which are widely distributed in the world. There are more than 100 species in my country. There are about 7 species that can be used as Scutellaria baicalensis, and Scutellaria baicalensis is one of them. The main medicinal component of this plant is baicalin, which has been proved by modern pharmacology to have the functions of clearing heat and detoxifying, stopping bleeding and preventing miscarriage, antibacterial and anti-inflammatory, protecting liver and gallbladder, reducing blood pressure and desensitization, sun protection and whitening. With the continuous incr...

Claims

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Application Information

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IPC IPC(8): C12N9/00C12N15/52C12N1/19C12N5/10
Inventor 孙敏雷桅
Owner SOUTHWEST UNIVERSITY
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