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Squalene synthase gene of Panax japonicus and application of the gene

A technology of squalene synthase and bamboo ginseng, which is applied in the fields of application, genetic engineering, plant genetic improvement, etc.

Inactive Publication Date: 2012-07-25
FUJIAN AGRI & FORESTRY UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there has been no literature report on the isolation and cloning of the full-length squalene synthase gene from the medicinal plant Panax ginseng

Method used

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  • Squalene synthase gene of Panax japonicus and application of the gene
  • Squalene synthase gene of Panax japonicus and application of the gene
  • Squalene synthase gene of Panax japonicus and application of the gene

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Embodiment 1

[0055] Embodiment 1 The cloning of bamboo ginseng squalene synthase gene

[0056] 1. tissue isolation

[0057] The bamboo-jointed ginseng plant comes from Jinggangshan, Jiangxi, and its roots are immediately frozen in liquid nitrogen and stored in a -80°C refrigerator.

[0058] 2. Total RNA isolation (RNA isolation)

[0059] Part of the tissue was taken and crushed with a mortar, added to a 1.5ml centrifuge tube filled with lysate, shaken thoroughly, and total RNA was extracted according to the instructions of the TRIzol reagent (TRIzol Reagents, Invitrogen, USA). The quality of total RNA was identified by agarose gel electrophoresis, and then the concentration and OD value of RNA were measured on an ultra-micro UV spectrophotometer.

[0060] 3. Cloning of the full-length cDNA of the target gene (Cloning of Full-length cDNA)

[0061] According to the conserved sequences of the squalene synthase genes that have been cloned from Arabidopsis thaliana, tobacco, licorice an...

Embodiment 2

[0073] Example 2 Sequence information and homology analysis of the squalene synthase gene of Bamboo ginseng

[0074] The full-length cDNA of the bamboo ginseng squalene synthase gene of the present invention is 1495bp in length, and the detailed sequence is shown in SEQ ID NO.1, wherein the open reading frame is located at nucleotides 87-1352. According to the full-length cDNA, the amino acid sequence of squalene synthase from bamboo ginseng was deduced, with a total of 421 amino acid residues, a molecular weight of 48.0kD, and a pI of 6.17. See SEQ ID NO.2 for the detailed sequence.

[0075] Log the obtained full-length cDNA sequence and the encoded amino acid sequence of the bamboo ginseng SQS gene on the NCBI website (http: / / www.ncbi.nlm.nih.gov), and use the BLAST program to search GenBank + EMBL + DDBJ respectively + PDB and non-redundant GenBank CDS translations + PDB + SwissProt + PIR + PRF database reported sequences, for nucleotide sequence and amino acid sequence h...

Embodiment 3

[0077] Example 3 Structural analysis and functional prediction of bamboo ginseng squalene synthase or polypeptide

[0078] The amino acid sequence of bamboo ginseng SQS was logged on the NCBI website, and a CDD (Conserved Domain Database) search was performed, and it was found that bamboo ginseng SQS had three conserved regions: 1'-1 trans isopentenyl diphosphate synthase (Trans_IPPS_HH), The enzyme catalyzes two isopentenyl diphosphate "head-to-head" condensation reactions; trans-isopentenyl diphosphate synthase (Trans_IPPS) and isoprenoid biosynthesis enzymes (Isoprenoid Biosynthesis enzymes), Belonging to the first family of isoprenoid biosynthetic enzymes, it can catalyze the synthesis of GPP, FPP or longer isoprenoid products. The catalytic site consists of a large central cavity consisting of two antiparallel α-double helices with walled aspartate-rich domains (DXXXD) that function primarily to bind prenyl phosphate groups , which can respectively catalyze the 1'-1 co...

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Abstract

The invention discloses a squalene synthase gene of Panax japonicus and application of the gene, and belongs to the technical field of biotechnology. The invention fills the blank that the squalene synthase gene has not been isolated or cloned from Panax japonicus as a traditional Chinese medicinal material. The squalene synthase gene provided by the invention has nucleotide sequences shown at 87th to 1352nd sites in SEQ ID No. 1 or homologous sequences in which one or more nucleotides are added, substituted, inserted or deleted, or alleles of the gene, or derivative nucleotide sequences. The squalene synthase gene provided by the invention can improve the content of saponin that is an essential active ingredient in ginseng by a genetic engineering technology, be applied to study and industrialization for improving the content of saponin via a transgenic technology, and facilitate quality improvement of ginseng medicinal material and relief of problem of deficient saponin medicinal resource, so that the gene has a good application prospect.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to squalene synthase gene expressed in bamboo ginseng and its application. Background technique [0002] Ginseng ( Panax ginseng C.A.Mey.) belongs to the Araliaceae ginseng plant and is one of the traditional precious medicinal materials. The ginseng plants have high medicinal value, such as enhancing immunity, anti-tumor, anti-arteriosclerosis and myocardial protection. The main functional components are ginsenosides, namely dammarane-type tetracyclic triterpenoids or balsamane-type pentacyclic triterpenoids. [0003] So far, ginsenosides have not been able to be synthesized artificially. Ginseng is a perennial herb with special cultivation and growth conditions and is very sensitive to soil and climate, making it difficult to promote and apply it on a large scale. [0004] Ginsenosides belong to the secondary products of the plant isoprenoid metabolic pathway - triterpe...

Claims

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Application Information

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IPC IPC(8): C12N15/54C12N9/10C12N15/63C12N1/21C12N1/19C12N5/10C12N15/82A01H5/00A01H5/06
Inventor 黄志伟许明郑金贵
Owner FUJIAN AGRI & FORESTRY UNIV
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